UI - 22422297 PMID- 12536225 DA - 20030121 IS - 0001-6322 VI - 105 IP - 2 DP - 2003 Feb TI - Distribution, progression and chemical composition of cortical amyloid-beta deposits in aged rhesus monkeys: similarities to the human. PG - 145-56 AB - A comprehensive investigation of the incidence, distribution, progression and chemical composition of Abeta deposits in the brains of two young (5 years) and seven aged (25-30 years) rhesus monkeys was conducted to determine the similarity of this phenomenon to that in the human. The brains of the young rhesus were devoid of Abeta deposits. In contrast, Abeta deposits were observed within the cerebral cortex of all aged animals. In animals with mild Abeta burden, deposits were observed primarily in association cortical zones. In animals with moderate Abeta burden, many paralimbic cortical zones also contained Abeta deposits. Finally, in an animal with a heavy burden of Abeta, core limbic cortical zones were also involved. The primary sensory and motor cortices were relatively free of Abeta deposits. A higher proportion of plaques contained Abeta40 as compared with Abeta42. Abeta deposits contained a number of other constituents. Cholinesterases were present in nearly 50% of plaques and displayed the exact same biochemical characteristics as those in the human. Nearly 20% of Abeta deposits also contained apolipoprotein E and a smaller proportion contained heparin sulfate proteoglycans and alpha1-anti-chymotrypsin. The latter three chemicals were present in many compact plaques. These results indicate that the distribution, progression and chemical composition of plaques in the aged rhesus monkey display many similarities to those observed in the aged human and Alzheimer's disease. Therefore, despite some differences from the human, the aged rhesus may be a good model for studies of the pathological effects of Abeta in the primate brain. AD - Laboratory for Neurodegenerative and Aging Research, Department of Medicine (Neuroscience), Harvard Medical School and Section of Gerontology, Beth Israel Deaconess Medical Center, 21-27 Burlington Ave., Boston, MA 02215, USA. FAU - Sani, Sepehr AU - Sani S FAU - Traul, David AU - Traul D FAU - Klink, Annelies AU - Klink A FAU - Niaraki, Nayson AU - Niaraki N FAU - Gonzalo-Ruiz, Alicia AU - Gonzalo-Ruiz A FAU - Wu, Chuang-Kuo AU - Wu CK FAU - Geula, Changiz AU - Geula C LA - eng PT - Journal Article CY - Germany TA - Acta Neuropathol (Berl) JID - 0412041 SB - IM EDAT- 2003/01/22 04:00 MHDA- 2003/01/22 04:00 PHST- 2002/Jul/13 [received] PHST- 2002/Sep/02 [revised] PHST- 2002/Sep/03 [accepted] PHST- 2002/Nov/05 [aheadofprint] AID - 10.1007/s00401-002-0626-5 [doi] PST - ppublish SO - Acta Neuropathol (Berl) 2003 Feb;105(2):145-56. UI - 0 PMID- 12214125 DA - 20020905 IS - 1387-2877 VI - 1 IP - 4,5 DP - 1999 Nov TI - Neurite-Outgrowth Regulating Functions of the Amyloid Protein Precursor of Alzheimer's Disease. PG - 275-285 AB - Many studies have shown that breakdown of the amyloid protein precursor (APP) to produce the amyloid protein is an important step in the pathogenic mechanism which causes Alzheimer's disease (AD). However, little is known about the normal function of APP. Developmental studies show that APP expression increases during the period of brain development when neurite outgrowth and synaptogenesis is maximal. APP is expressed highly within growing neurites and in growth cones, and purified APP has been shown to stimulate neurite outgrowth from cells in culture. Thus APP may regulate neurite outgrowth or synaptogenesis in vivo. APP is actively secreted from many cells, and the C-terminally secreted APP has been shown to associate with components of the extracellular matrix, such as the heparan sulphate proteoglycans (HSPGs). Two putative heparin-binding domains on APP have been reported. Binding of HSPGs to an N-terminal heparin-binding domain (HBD-1) stimulates the effect of substrate-bound APP on neurite outgrowth. In the mature nervous system, APP may play an important role in the regulation of wound repair. It is highly likely that studies on the normal functions of APP will shed further light on aspects of the pathogenesis of AD. AD - Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia. AU - Small DH AU - Clarris HL AU - Williamson TG AU - Reed G AU - Key B AU - Mok SS AU - Beyreuther K AU - Masters CL AU - Nurcombe V LA - ENG PT - JOURNAL ARTICLE TA - J Alzheimers Dis JID - 9814863 EDAT- 2002/09/06 10:00 MHDA- 2002/09/06 10:00 PST - ppublish SO - J Alzheimers Dis 1999 Nov;1(4,5):275-285. UI - 22095352 PMID- 12100726 DA - 20020708 DCOM- 20020828 IS - 0019-2805 VI - 106 IP - 3 DP - 2002 Jul TI - Effects of C-reactive protein and pentosan polysulphate on human complement activation. PG - 381-8 AB - Complement (C) activation is believed to play an adverse role in several chronic degenerative disease processes, including atherosclerosis, myocardial infarction and Alzheimer's disease. We developed several in vitro quantitative assays to evaluate processes which activate C in human serum, and to assess candidates which might block that activation. Binding of C-reactive protein (CRP) to immobilized cell surfaces was used as a tissue-based method of activation, while immunoglobulin G in solution was used as a surrogate antibody method. Activation was assessed by deposition of C fragments on fixed cell surfaces, or by capture of C5b-9 from solution. We observed that several cell lines, including SH-SY5Y, U-937, THP-1 and ECV304, bound CRP and activated C following attachment of cells to a plastic surface by means of air drying. Treatment of human neuroblastoma SH-SY5Y cells with the reactive oxygen intermediates generated by xanthine (Xa) - xanthine oxidase (XaOx) prior to air drying or by hydrogen peroxide solutions after air drying, enhanced C activation, possibly through oxidation of the cell lipid membrane. Several C inhibitors were tested for their effectiveness in blocking these systems. Pentosan polysulphate (PPS), an orally active agent, blocked C activation in the same concentration range of 1-1000 microg/ml as heparin, dextran sulphate, compstatin and fucoidan. PPS may have practical application as a C inhibitor. AD - Kinsmen Laboratory of Neurological Research, University of British Columbia, Vancouver, Canada. FAU - Klegeris, Andis AU - Klegeris A FAU - Singh, Edith A AU - Singh EA FAU - McGeer, Patrick L AU - McGeer PL LA - eng PT - Journal Article CY - England TA - Immunology JID - 0374672 RN - 0 (Complement Inactivators) RN - 0 (Complement Membrane Attack Complex) RN - 37300-21-3 (Pentosan Sulfuric Polyester) RN - 9007-41-4 (C-Reactive Protein) SB - IM MH - C-Reactive Protein/*pharmacology MH - Complement Activation/*drug effects MH - Complement Inactivators/pharmacology MH - Complement Membrane Attack Complex/immunology MH - Human MH - Pentosan Sulfuric Polyester/*pharmacology MH - Support, Non-U.S. Gov't MH - Tumor Cells, Cultured EDAT- 2002/07/09 10:00 MHDA- 2002/08/29 10:01 AID - 1425 [pii] PST - ppublish SO - Immunology 2002 Jul;106(3):381-8. UI - 22194392 PMID- 12095987 DA - 20020902 DCOM- 20021029 LR - 20030103 IS - 0021-9258 VI - 277 IP - 36 DP - 2002 Sep 6 TI - Fibroblast growth factor 1 regulates signaling via the glycogen synthase kinase-3beta pathway. Implications for neuroprotection. PG - 32985-91 AB - We hypothesize that in neurodegenerative disorders such as Alzheimer's disease and human immunodeficiency virus encephalitis the neuroprotective activity of fibroblast growth factor 1 (FGF1) against several neurotoxic agents might involve regulation of glycogen synthase kinase-3beta (GSK3beta), a pathway important in determining cell fate. In primary rat neuronal and HT22 cells, FGF1 promoted a time-dependent inactivation of GSK3beta by phosphorylation at serine 9. Blocking FGF1 receptors with heparinase reduced this effect. The effects of FGF1 on GSK3beta were dependent on phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) because inhibitors of this pathway or infection with dominant negative Akt adenovirus blocked inactivation. Furthermore, treatment of neuronal cells with FGF1 resulted in ERK-independent Akt phosphorylation and beta-catenin translocation into the nucleus. On the other hand, infection with wild-type GSK3beta recombinant adenovirus-associated virus increased activity of GSK3beta and cell death, both of which were reduced by FGF1 treatment. Moreover, FGF1 protection against glutamate toxicity was dependent on GSK3beta inactivation by the PI3K-Akt but was independent of ERK. Taken together these results suggest that neuroprotective effects of FGF1 might involve inactivation of GSK3beta by a pathway involving activation of the PI3K-Akt cascades. AD - Department of Neurosciences, University of California San Diego, La Jolla, California 92093-0624, USA. FAU - Hashimoto, Makoto AU - Hashimoto M FAU - Sagara, Yutaka AU - Sagara Y FAU - Langford, Dianne AU - Langford D FAU - Everall, Ian P AU - Everall IP FAU - Mallory, Margaret AU - Mallory M FAU - Everson, Analisa AU - Everson A FAU - Digicaylioglu, Murat AU - Digicaylioglu M FAU - Masliah, Eliezer AU - Masliah E LA - eng ID - AG01029/AG/NIA ID - DA12065/DA/NIDA ID - MH45294/MH/NIMH ID - MH58164/MH/NIMH ID - MH59745/MH/NIMH ID - MH62963/MH/NIMH PT - Journal Article CY - United States TA - J Biol Chem JID - 2985121R RN - 0 (Cytoskeletal Proteins) RN - 0 (Genetic Vectors) RN - 0 (Neuroprotective Agents) RN - 0 (Proto-Oncogene Proteins) RN - 0 (Recombinant Proteins) RN - 0 (Trans-Activators) RN - 0 (proto-oncogene protein akt) RN - 104781-85-3 (Fibroblast Growth Factor 1) RN - 146409-33-8 (beta catenin) RN - 56-45-1 (Serine) RN - 56-86-0 (Glutamic Acid) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.137 (1-Phosphatidylinositol 3-Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) RN - EC 4.2.2.7 (Heparin Lyase) SB - IM MH - 1-Phosphatidylinositol 3-Kinase/metabolism MH - Adenoviridae/metabolism MH - Animal MH - Blotting, Western MH - Ca(2+)-Calmodulin Dependent Protein Kinase/*metabolism MH - Cell Death MH - Cell Line MH - Cell Survival MH - Cells, Cultured MH - Cytoskeletal Proteins/metabolism MH - DNA Fragmentation MH - Fibroblast Growth Factor 1/metabolism/*pharmacology MH - Genetic Vectors MH - Glutamic Acid/metabolism MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Heparin Lyase/metabolism MH - Human MH - Immunohistochemistry MH - Microscopy, Confocal MH - Microscopy, Fluorescence MH - Models, Biological MH - Neurons/metabolism MH - Neuroprotective Agents/*pharmacology MH - Phosphorylation MH - Proto-Oncogene Proteins/metabolism MH - Rats MH - Recombinant Proteins/metabolism MH - Serine/metabolism MH - *Signal Transduction MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Time Factors MH - Trans-Activators/metabolism EDAT- 2002/07/04 10:00 MHDA- 2002/10/31 04:00 PHST- 2002/Jul/02 [aheadofprint] AID - 10.1074/jbc.M202803200 [doi] AID - M202803200 [pii] PST - ppublish SO - J Biol Chem 2002 Sep 6;277(36):32985-91. UI - 22074875 PMID- 12079686 DA - 20020624 DCOM- 20030103 IS - 0014-2999 VI - 445 IP - 3 DP - 2002 Jun 12 TI - Low molecular weight glycosaminoglycan blockade of beta-amyloid induced neuropathology. PG - 211-20 AB - Previous studies have shown different roles for proteoglycans and glycosaminoglycans (GAGs) in Alzheimer's disease (AD) neuropathology. Using a rat model of beta-amyloid induced neuropathology, we tested whether low molecular weight glycosaminoglycans (Certoparin and C6) could be useful as preventative agents and/or as a potential therapeutic treatment for AD. Chronic subcutaneous low molecular weight glycosaminoglycan injections beginning either before or after an intra-amygdaloid beta-amyloid-(25-35) injection blocked abnormal intracellular tau changes and reactive astrocytosis but did not affect beta-amyloid's aggregation state. Also, low molecular weight glycosaminoglycan injections beginning 1 day prior to sacrifice did not block the effects of beta-amyloid nor did injections of a disaccharide, suggesting chronic low molecular weight glycosaminoglycan treatment is needed to block the effects of beta-amyloid. Furthermore, these data indicate that there is a molecular weight range of active low molecular weight glycosaminoglycans in this model; and supports the investigation of low molecular weight glycosaminoglycans as a preventative and/or therapeutic treatment of beta-amyloid induced neuropathology. AD - Department of Pharmacology, Loyola University Chicago Medical Center, 2160 South First Avenue, Rm. 2638, Maywood, IL 60153, USA. FAU - Walzer, Mark AU - Walzer M FAU - Lorens, Stanley AU - Lorens S FAU - Hejna, Matthew AU - Hejna M FAU - Fareed, Jawed AU - Fareed J FAU - Hanin, Israel AU - Hanin I FAU - Cornelli, Umberto AU - Cornelli U FAU - Lee, John M AU - Lee JM LA - eng PT - Journal Article CY - Netherlands TA - Eur J Pharmacol JID - 1254354 RN - 0 (Amyloid beta-Protein) RN - 0 (Glycosaminoglycans) RN - 0 (Heparin, Low-Molecular-Weight) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta-protein (25-35)) RN - 0 (certoparin) RN - 0 (tau Proteins) SB - IM MH - Amyloid beta-Protein/analysis/*metabolism/pharmacology MH - Animal MH - Astrocytes/chemistry/drug effects/pathology MH - Glycosaminoglycans/pharmacology MH - Heparin, Low-Molecular-Weight/*pharmacology MH - Hippocampus/chemistry/*drug effects/pathology MH - Male MH - Peptide Fragments/analysis/*metabolism/pharmacology MH - Rats MH - Rats, Inbred F344 MH - Support, Non-U.S. Gov't MH - tau Proteins/analysis/metabolism EDAT- 2002/06/25 10:00 MHDA- 2003/01/04 04:00 AID - S0014299902017594 [pii] PST - ppublish SO - Eur J Pharmacol 2002 Jun 12;445(3):211-20. UI - 22064987 PMID- 12069613 DA - 20020618 DCOM- 20020712 IS - 0006-2960 VI - 41 IP - 25 DP - 2002 Jun 25 TI - Inhibition of apolipoprotein E-related neurotoxicity by glycosaminoglycans and their oligosaccharides. PG - 8203-11 AB - Apolipoprotein E (apoE) has been genetically linked to late-onset Alzheimer's disease (AD). The role of this lipid-transport protein in AD remains to be established. One hypothesis is that apoE, particularly the apoE4 isoform, may have neurotoxic effects as demonstrated using apoE-related synthetic peptides and the N-terminal fragment of apoE. ApoE is a heparan-sulfate binding protein, and apoE peptide neurotoxicity can be blocked by heparin and prevented by degrading heparan sulfate or inhibiting its biosynthesis. The possibility that heparin inhibition of toxicity is mediated by a specific oligosaccharide sequence was investigated using a bioassay to determine the inhibition of apoE peptide toxicity by glycosaminoglycans and purified glycosaminoglycan oligosaccharides. Studies on modified heparins showed that the presence of N-sulfo groups and either 2- or 6-O sulfo groups were required for inhibition of toxicity. Heparin oligosaccharides with eight or more saccharide residues with seven O-sulfo groups and four N-sulfo groups exhibited potent inhibition. Larger oligosaccharides, and heparin and heparan sulfate polymers, afforded comparable, or somewhat better, protective effects but also caused clumping and detachment of cells when administrated alone. AD - ApoLogic, Inc., Cincinnati, Ohio, USA. FAU - Bazin, Helene G AU - Bazin HG FAU - Marques, Marcos A AU - Marques MA FAU - Owens, Albert P 3rd AU - Owens AP 3rd FAU - Linhardt, Robert J AU - Linhardt RJ FAU - Crutcher, Keith A AU - Crutcher KA LA - eng ID - GM 38060/GM/NIGMS ID - HL 52622/HL/NHLBI ID - NS 037986/NS/NINDS PT - Journal Article CY - United States TA - Biochemistry JID - 0370623 RN - 0 (Apolipoproteins E) RN - 0 (Glycosaminoglycans) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Heparin, Low-Molecular-Weight) RN - 0 (Oligosaccharides) RN - 24967-94-0 (Dermatan Sulfate) RN - 9005-49-6 (Heparin) SB - IM MH - Animal MH - Apolipoproteins E/*antagonists & inhibitors/metabolism/*toxicity MH - Binding, Competitive/drug effects MH - Cell Death/drug effects MH - Cells, Cultured MH - Chick Embryo MH - Dermatan Sulfate/metabolism/pharmacology MH - Ganglia, Sympathetic/cytology/drug effects/metabolism MH - Glycosaminoglycans/metabolism/*pharmacology MH - Heparan Sulfate Proteoglycan/metabolism/pharmacology MH - Heparin/metabolism/pharmacology MH - Heparin, Low-Molecular-Weight/metabolism/pharmacology MH - Neurons/*drug effects/metabolism MH - Oligosaccharides/metabolism/*pharmacology MH - Partial Thromboplastin Time MH - Protein Binding/drug effects MH - Support, U.S. Gov't, P.H.S. MH - Swine EDAT- 2002/06/19 10:00 MHDA- 2002/07/13 10:01 AID - bi025817e [pii] PST - ppublish SO - Biochemistry 2002 Jun 25;41(25):8203-11. UI - 22058862 PMID- 12062547 DA - 20020613 IS - 0049-3848 VI - 105 IP - 5 DP - 2002 Mar 1 TI - The blood-brain barrier accessibility of a heparin-derived oligosaccharides C3. PG - 447-53 AB - Although heparin-derived oligosaccharide(s) (HDO) have been clinically used for the management of neurological disorders, such as stroke and Alzheimer's disease (AD), very little information on the mechanism of their therapeutic action is known. To test the hypothesis that HDO may pass through the blood-brain barrier (BBB) to mediate their effects, a pharmacodynamic (PD) model was developed and the presence of HDO in the cerebrospinal fluid (CSF) was used as a BBB accessibility index. Rats were treated with an ultralow molecular weight (MW) heparin fragment C3 via the intravenous or subcutaneous routes at 5-10 mg/kg. At varying periods, the plasma, CSF, and brain samples were collected, and functional anti-factor Xa activities were measured to quantitate the CSF/plasma ratios (CPR) and the brain uptake. C3 showed CPR of 1.7% and 0.8% after intravenous and subcutaneous injections, respectively. These findings were verified by intravenous administration of tritium-labeled C3 followed by detection of the radioactivity in the CSF and brain homogenates. These data suggest that ultralow MW HDO may pass through the BBB. AD - Department of Pharmacology and Experimental Therapeutics, Stritch School of Medicine, Loyola University Chicago, 2160 South First Avenue, Maywood, IL 60153, USA. qma1@luc.edu FAU - Ma, Qing AU - Ma Q FAU - Dudas, Bertalan AU - Dudas B FAU - Hejna, Matthew AU - Hejna M FAU - Cornelli, Umberto AU - Cornelli U FAU - Lee, John M AU - Lee JM FAU - Lorens, Stanley AU - Lorens S FAU - Mervis, Ronald AU - Mervis R FAU - Hanin, Israel AU - Hanin I FAU - Fareed, Jawed AU - Fareed J LA - eng ID - 1R41 AG15740-01/AG/NIA PT - Journal Article CY - United States TA - Thromb Res JID - 0326377 SB - IM EDAT- 2002/06/14 10:00 MHDA- 2002/06/14 10:00 AID - S0049384802000506 [pii] PST - ppublish SO - Thromb Res 2002 Mar 1;105(5):447-53. UI - 22057385 PMID- 12062474 DA - 20020613 DCOM- 20020807 IS - 0006-8993 VI - 935 IP - 1-2 DP - 2002 May 10 TI - Sulfo-glycosaminoglycan content affects PHF-tau solubility and allows the identification of different types of PHFs. PG - 65-72 AB - Sulfo-glycosaminoglycans (sGAGs) are involved in the assembly of tau in at least a subpopulation of paired helical filaments (PHFs) in Alzheimer's disease (AD). To further understand the role of sGAG molecules in the structure of PHFs, we isolated PHFs from patients with AD and treated them with heparinase. Immunoelectron microscopy and Western blotting (WB) were used later on to analyze the changes obtained. The heparinase treatment abolished Tau14 and AT8 immunodecoration (two N-terminal tau antibodies) and increased PHF-1 labeling (a C-terminal antibody). In addition, heparinase-treated filaments are more labile than control ones as demonstrated by sodium dodecyl sulfate-extraction and subsequent WB. In summary, our results demonstrate that sGAG content affects PHF conformation as well as PHF-tau solubilization. AD - Severo Ochoa Center for Molecular Biology, CSIC/UAM, Faculty of Sciences, Universidad Autonoma de Madrid, Cantoblanco, 28049 Madrid, Spain. FAU - Hernandez, Felix AU - Hernandez F FAU - Perez, Mar AU - Perez M FAU - Lucas, Jose J AU - Lucas JJ FAU - Avila, Jesus AU - Avila J LA - eng PT - Journal Article CY - Netherlands TA - Brain Res JID - 0045503 RN - 0 (Antibodies) RN - 0 (Epitopes) RN - 0 (Glycosaminoglycans) RN - 0 (Protein Isoforms) RN - 0 (Sulfuric Acid Esters) RN - 0 (tau Proteins) RN - EC 4.2.2.7 (Heparin Lyase) SB - IM MH - Alzheimer Disease/*metabolism/pathology/physiopathology MH - Amino Acid Sequence/physiology MH - Antibodies/diagnostic use MH - Brain/*metabolism/pathology/ultrastructure MH - Epitopes/chemistry/immunology MH - Glycosaminoglycans/*chemistry MH - Heparin Lyase/diagnostic use MH - Human MH - Immunohistochemistry MH - Microscopy, Electron MH - Molecular Conformation MH - Neurofibrillary Tangles/*chemistry/ultrastructure MH - Neurons/*metabolism/pathology/ultrastructure MH - Protein Isoforms/chemistry/ultrastructure MH - Protein Structure, Tertiary/drug effects/physiology MH - Solubility MH - Sulfuric Acid Esters/*chemistry MH - Support, Non-U.S. Gov't MH - tau Proteins/*chemistry/ultrastructure EDAT- 2002/06/14 10:00 MHDA- 2002/08/08 10:01 AID - S0006899302024551 [pii] PST - ppublish SO - Brain Res 2002 May 10;935(1-2):65-72. UI - 22029039 PMID- 12031824 DA - 20020528 DCOM- 20021204 IS - 0049-3848 VI - 105 IP - 4 DP - 2002 Feb 15 TI - Molecular and biochemical profiling of a heparin-derived oligosaccharide, C3. PG - 303-9 AB - This study was designed to characterize a heparin-derived oligosaccharide (HDO), C3, using chemical and biochemical methods. Although previous studies have suggested C3 as a promising compound in the treatment of Alzheimer's disease (AD), its molecular and biochemical properties are still unknown. In this study, the molecular profiles and anticoagulant effects of C3 were investigated. To characterize the molecular and biochemical properties of C3, gel permeation chromatography (GPC), polyacrylmide gel electrophoresis (PAGE), radiolabeling and anticoagulant assays, such as activated partial thromboplastin time (APTT), Heptest, and anti-factor Xa assay, were used. The GPC profile revealed that C3 was an ultra-low-molecular-weight (MW) heparin mixture. The multiple components in C3 were studied with PAGE analysis. Tritium-labeled C3 exhibited similar biological properties as nonlabeled materials. The biological assays showed that C3 and its components exhibited weak anticoagulant effect. These results demonstrated the applicability of the combination of GPC, PAGE, and coagulation assays to characterize the molecular and biochemical profile of HDO. In addition, the low anticoagulant effect of C3 suggests that this compound could be a relatively low-risk adjunct in the treatment of AD. AD - Department of Pharmacology, Stritch School of Medicine, Loyola University Chicago, Maywood, IL 60153, USA. FAU - Ma, Qing AU - Ma Q FAU - Dudas, Bertalan AU - Dudas B FAU - Daud, Asif AU - Daud A FAU - Iqbal, Omer AU - Iqbal O FAU - Hoppensteadt, Debra AU - Hoppensteadt D FAU - Jeske, Walter AU - Jeske W FAU - Cornelli, Umberto AU - Cornelli U FAU - Lee, John AU - Lee J FAU - Lorens, Stanley AU - Lorens S FAU - Mervis, Ronald AU - Mervis R FAU - Hanin, Israel AU - Hanin I FAU - Capila, Ishan AU - Capila I FAU - Linhardt, Robert AU - Linhardt R FAU - Fareed, Jawed AU - Fareed J LA - eng ID - 1R41 AG15740-01/AG/NIA PT - Journal Article CY - United States TA - Thromb Res JID - 0326377 RN - 0 (Anticoagulants) RN - 0 (Oligosaccharides) RN - 10028-17-8 (Tritium) RN - 9005-49-6 (Heparin) RN - EC 3.4.21.6 (Factor Xa) SB - IM MH - Animal MH - Anticoagulants/chemistry/isolation & purification/pharmacology MH - Blood Coagulation Tests MH - Electrophoresis, Polyacrylamide Gel MH - Factor Xa/antagonists & inhibitors MH - Heparin/*chemistry/isolation & purification/pharmacology MH - Human MH - In Vitro MH - Molecular Weight MH - Oligosaccharides/*chemistry/isolation & purification/pharmacology MH - Partial Thromboplastin Time MH - Support, U.S. Gov't, P.H.S. MH - Tritium EDAT- 2002/05/29 10:00 MHDA- 2002/12/05 04:00 AID - S0049384801004133 [pii] PST - ppublish SO - Thromb Res 2002 Feb 15;105(4):303-9. UI - 22007775 PMID- 12009502 DA - 20020515 DCOM- 20020708 LR - 20021213 IS - 0197-4580 VI - 23 IP - 4 DP - 2002 Jul-Aug TI - Heparin attenuates cytotoxic and inflammatory activity of Alzheimer amyloid-beta in vitro. PG - 531-6 AB - Amyloid-beta protein (Abeta) is implicated in the pathogenesis of Alzheimer's disease because of its neurotoxicity and the ability to trigger local inflammation. Compounds that interact with the amino acids of the N-terminal region or interfere with aggregation can reduce the Abeta biologic activity. We evaluated the effect of heparin on Abeta (1-42) neurotoxicity and on its ability to activate complement and contact system. On differentiated PC12 cells, a reliable model of neuronal cells, heparin at the doses of 10 and 20 microg/ml significantly counteracted Abeta cytotoxicity as assessed by measuring MTT conversion. We then explored the effect of heparin on Abeta (1-42)-induced complement and contact system activation. Abeta (1-42) was incubated with heparin in presence of normal plasma as the source of complement and contact system factors. Heparin reduced, in a dose-dependent manner, complement and contact system activation, assessed by measuring the degree of C4 and high molecular weight kininogen cleavage. The present data show that heparin can attenuate neurotoxic and pro-inflammatory activity of Abeta and suggest that this drug could represent a new strategy to reduce the progressive neurodegeneration in AD. AD - Department of Internal Medicine, Ospedale Maggiore IRCCS, University of Milan, Italy. Luigi.Bergamaschini@unimi.it FAU - Bergamaschini, Luigi AU - Bergamaschini L FAU - Donarini, Cesare AU - Donarini C FAU - Rossi, Emanuela AU - Rossi E FAU - De Luigi, Ada AU - De Luigi A FAU - Vergani, Carlo AU - Vergani C FAU - De Simoni, Maria Grazia AU - De Simoni MG LA - eng PT - Journal Article CY - United States TA - Neurobiol Aging JID - 8100437 RN - 0 (Amyloid beta-Protein) RN - 0 (Anticoagulants) RN - 0 (Indicators and Reagents) RN - 0 (Peptide Fragments) RN - 0 (beta-amyloid (1-42)) RN - 9005-49-6 (Heparin) SB - IM MH - Amyloid beta-Protein/*antagonists & inhibitors/isolation & purification/*toxicity MH - Animal MH - Anticoagulants/*pharmacology MH - Blotting, Western MH - Cell Survival/drug effects MH - Cells, Cultured MH - Complement Activation/drug effects MH - Densitometry MH - Heparin/*pharmacology MH - Human MH - Indicators and Reagents MH - Inflammation/chemically induced/*prevention & control MH - Neurons/drug effects MH - PC12 Cells MH - Peptide Fragments/antagonists & inhibitors/isolation & purification/toxicity MH - Rats MH - Support, Non-U.S. Gov't EDAT- 2002/05/16 10:00 MHDA- 2002/07/09 10:01 AID - S0197458002000039 [pii] PST - ppublish SO - Neurobiol Aging 2002 Jul-Aug;23(4):531-6. UI - 21630114 PMID- 11755024 DA - 20011228 DCOM- 20020228 IS - 0197-4580 VI - 23 IP - 1 DP - 2002 Jan-Feb TI - Oral and subcutaneous administration of the glycosaminoglycan C3 attenuates Abeta(25-35)-induced abnormal tau protein immunoreactivity in rat brain. PG - 97-104 AB - High molecular weight glycosaminoglycans (GAG) and proteoglycans (PG) affect pathological changes of the brain in Alzheimer's disease (AD). PG stimulate the processing and aggregation of amyloid-beta (Abeta), protect the protein from proteolysis, and increase the formation of neurofibrillary tangles by inducing the hyperphosphorylation of tau protein. These effects may be competitively inhibited by GAG.We have studied the effects of orally (by gavage) and subcutaneously (s.c.) administered low molecular weight heparin, C3 (4-10 oligosaccharides; MW = 2.1 kDa; USP value = 12 U/mg), on abnormal tau-2 protein immunoreactivity in the rat hippocampus following a single, unilateral intra-amygdaloid administration of Abeta(25-35). Oral administration of C3 (25 mg/kg; once daily) was initiated 3 days prior to Abeta(25-35) administration, and was continued daily for an additional 14 days. S.c. administration of C3 (2.5 mg/kg, twice daily), was started 3 days prior to, and was continued for 32 days after, Abeta(25-35) administration. Animal brains were subsequently processed for tau-2, ChAT-immunoreactivity, choline acetyltransferase (ChAT) activity and acetylcholinesterase (AChE) activity. Both oral and s.c. administration of C3 attenuated Abeta(25-35) induced appearance of tau-2-immunoreactive (IR) perikarya in the ipsilateral hippocampus (P < 0.05). Hippocampal cholinergic enzyme activity in C3 treated animals was not significantly different from control animals.The present findings suggest that C3 might be used successfully to prevent abnormal tau protein formation in chronic neurologic diseases, such as AD. Moreover, our data demonstrate that the mechanism of this effect does not appear to influence the cholinergic system of the brain. AD - Loyola University Chicago, Stritch School of Medicine, Columbus, Ohio 43212, USA. bdudas@luc.edu FAU - Dudas, B AU - Dudas B FAU - Cornelli, U AU - Cornelli U FAU - Lee, J M AU - Lee JM FAU - Hejna, M J AU - Hejna MJ FAU - Walzer, M AU - Walzer M FAU - Lorens, S A AU - Lorens SA FAU - Mervis, R F AU - Mervis RF FAU - Fareed, J AU - Fareed J FAU - Hanin, I AU - Hanin I LA - eng ID - 1 R4 AG15740-01/AG/NIA PT - Journal Article CY - United States TA - Neurobiol Aging JID - 8100437 RN - 0 (Amyloid beta-Protein) RN - 0 (Heparin, Low-Molecular-Weight) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta-protein (25-35)) RN - 0 (tau Proteins) RN - EC 2.3.1.6 (Choline O-Acetyltransferase) RN - EC 3.1.1.7 (Acetylcholinesterase) SB - IM MH - Acetylcholinesterase/metabolism MH - Administration, Oral MH - Amyloid beta-Protein/administration & dosage/*pharmacology MH - Animal MH - Brain Chemistry/*drug effects MH - Choline O-Acetyltransferase/metabolism MH - Heparin, Low-Molecular-Weight/administration & dosage/*pharmacology MH - Hippocampus/drug effects/enzymology/metabolism MH - Immunohistochemistry MH - Injections MH - Injections, Subcutaneous MH - Male MH - Peptide Fragments/administration & dosage/*pharmacology MH - Rats MH - Rats, Inbred F344 MH - Stereotaxic Techniques MH - Support, U.S. Gov't, P.H.S. MH - tau Proteins/*metabolism EDAT- 2002/01/05 10:00 MHDA- 2002/03/01 10:01 AID - S019745800100255X [pii] PST - ppublish SO - Neurobiol Aging 2002 Jan-Feb;23(1):97-104. UI - 21487111 PMID- 11601501 DA - 20011016 DCOM- 20011101 IS - 0364-5134 VI - 50 IP - 4 DP - 2001 Oct TI - Pick's disease associated with the novel Tau gene mutation K369I. PG - 503-13 AB - Exonic and intronic mutations in Tau cause neurodegenerative syndromes characterized by frontotemporal dementia and filamentous tau protein deposits. We describe a K369I missense mutation in exon 12 of Tau in a patient with a pathology typical of sporadic Pick's disease. The proband presented with severe personality changes, followed by loss of cognitive function. Detailed postmortem examination of the brain showed atrophy, which was most pronounced in the temporal lobes; and numerous tau-immunoreactive Pick bodies and Pick cells in the neocortex and the hippocampal formation, as well as in subcortical brain regions. Their appearance and staining characteristics were indistinguishable from those of sporadic Pick's disease. However, immunoblot analysis of sarkosyl-insoluble tau showed three major bands of 60, 64, and 68 kDa, consistent with the presence of 3- and 4-repeat tau isoforms, as in Alzheimer's disease. Isolated tau filaments were irregularly twisted ribbons, with a small number of Alzheimer-type paired helical filaments. In the presence of heparin, tau proteins with the K369I mutation formed short, slender filaments. Biochemically, recombinant tau proteins with the K369I mutation showed reduced ability to promote microtubule assembly, suggesting that this may be the primary effect of the mutation by providing a pool of aberrant tau for filament assembly. Taken together, results indicate that the K369I mutation in Tau can cause a dementing disease with a neuropathology like that of Pick's disease. AD - Institute of Neuropathology, Ludwig-Maximilians-University, Munich, Germany. FAU - Neumann, M AU - Neumann M FAU - Schulz-Schaeffer, W AU - Schulz-Schaeffer W FAU - Crowther, R A AU - Crowther RA FAU - Smith, M J AU - Smith MJ FAU - Spillantini, M G AU - Spillantini MG FAU - Goedert, M AU - Goedert M FAU - Kretzschmar, H A AU - Kretzschmar HA LA - eng PT - Journal Article CY - United States TA - Ann Neurol JID - 7707449 RN - 0 (tau Proteins) SB - IM MH - DNA Mutational Analysis MH - Female MH - Frontal Lobe/pathology MH - Human MH - Immunoblotting MH - Immunohistochemistry MH - Lateral Ventricles/pathology MH - Microscopy, Electron MH - Microtubules/metabolism MH - Middle Age MH - *Mutation, Missense MH - Pick Disease of the Brain/*genetics/pathology MH - Temporal Lobe/pathology MH - tau Proteins/*genetics/metabolism/ultrastructure EDAT- 2001/10/17 10:00 MHDA- 2001/11/03 10:01 PST - ppublish SO - Ann Neurol 2001 Oct;50(4):503-13. UI - 21611209 PMID- 11600492 DA - 20011217 DCOM- 20020131 LR - 20030103 IS - 0021-9258 VI - 276 IP - 51 DP - 2001 Dec 21 TI - Metalloprotease-dependent protransforming growth factor-alpha ectodomain shedding in the absence of tumor necrosis factor-alpha-converting enzyme. PG - 48510-7 AB - Zinc-dependent metalloproteases can mediate the shedding of the extracellular domain of many unrelated transmembrane proteins from the cell surface. In most instances, this process, also known as ectodomain shedding, is regulated via protein kinase C (PKC). The tumor necrosis factor alpha-converting enzyme (TACE) was the first protease involved in regulated protein ectodomain shedding identified. Although TACE belongs to the family of metalloprotease-disintegrins, few members of this family have been shown to participate in regulated ectodomain shedding. In fact, the phenotype of tace-/- cells and that of Chinese hamster ovary cell mutants defective in ectodomain shedding points to the existence of a common PKC-activated ectodomain shedding system, whose proteolytic component is TACE, that acts on a variety of transmembrane proteins. Examples of these proteins include the Alzheimer's disease-related protein beta-amyloid precursor protein (betaAPP) and the transmembrane growth factors protransforming growth factor-alpha (pro-TGF-alpha) and, as shown in this report, proheparin-binding epidermal growth factor-like growth factor (pro-HB-EGF). Here we show that the mercurial compound 4-aminophenylmercuric acetate (APMA), frequently used to activate in vitro recombinant matrix metalloproteases, is an activator of the shedding of betaAPP, pro-HB-EGF, and pro-TGF-alpha. Treatment of tace-/- cells or Chinese hamster ovary shedding-defective mutants with APMA activates the cleavage of pro-TGF-alpha but not that of pro-HB-EGF or betaAPP, indicating that APMA activates TACE and also a previously unacknowledged proteolytic activity specific for pro-TGF-alpha. Characterization of this proteolytic activity indicates that it acts on pro-TGF-alpha located at the cell surface and that it is a metalloprotease active in cells defective in furin activity. In summary, treatment of shedding-defective cell lines with APMA unveils the existence of a metalloprotease activity alternative to TACE with the ability to specifically shed the ectodomain of pro-TGF-alpha. AD - Laboratori de Recerca Oncologica, Servei d'Oncologia Medica, Hospital Universitari Vall d'Hebron, Psg. Vall d'Hebron 119-129, Barcelona 08035, Spain. FAU - Merlos-Suarez, A AU - Merlos-Suarez A FAU - Ruiz-Paz, S AU - Ruiz-Paz S FAU - Baselga, J AU - Baselga J FAU - Arribas, J AU - Arribas J LA - eng PT - Journal Article CY - United States TA - J Biol Chem JID - 2985121R RN - 0 (Membrane Proteins) RN - 0 (Transforming Growth Factor alpha) RN - 149176-25-0 (heparin-binding EGF-like growth factor) RN - 62-38-4 (Phenylmercuric Acetate) RN - 62229-50-9 (Epidermal Growth Factor) RN - 6283-24-5 (4-aminophenylmercuriacetate) RN - EC 2.7.1.37 (Protein Kinase C) RN - EC 3.4.24 (Metalloendopeptidases) RN - EC 3.4.24.- (TNF-alpha converting enzyme) SB - IM MH - Animal MH - CHO Cells MH - Cell Line MH - Epidermal Growth Factor/metabolism MH - Hamsters MH - Human MH - Hydrolysis MH - Membrane Proteins/metabolism MH - Metalloendopeptidases/genetics/*metabolism/*physiology MH - Phenylmercuric Acetate/*analogs & derivatives/pharmacology MH - Protein Kinase C/metabolism MH - Support, Non-U.S. Gov't MH - Transforming Growth Factor alpha/*metabolism EDAT- 2001/10/16 10:00 MHDA- 2002/02/01 10:01 PHST- 2001/Oct/12 [aheadofprint] AID - 10.1074/jbc.M103488200 [doi] AID - M103488200 [pii] PST - ppublish SO - J Biol Chem 2001 Dec 21;276(51):48510-7. UI - 21475974 PMID- 11591887 DA - 20011009 DCOM- 20020214 IS - 1076-1551 VI - 7 IP - 8 DP - 2001 Aug TI - Inhibition of amyloidosis using low-molecular-weight heparins. PG - 517-22 AB - BACKGROUND: Amyloid diseases are characterized by the tissue deposition of extracellular proteinaceous material, which results in organ dysfunction and death. Colocalization of heparan sulfate (HS) proteoglycans to amyloid deposits suggests that they may be an early event in amyloid formation and play an important role in fibril formation. Structural analysis has demonstrated that HS interacts with amyloidogenic proteins resulting in structural changes that allow for an increase in beta-sheet content, possibly enhancing fibrillogenesis. Recent studies have shown that small-molecule anionic sulfonates or sulfates can arrest inflammation-associated (AA) amyloid induction. MATERIALS AND METHODS: In the present study, we examine the effect of low-molecular-weight heparins (LMWHs) on the development of amyloid in the mouse model of AA amyloid. In addition, in vitro fibril formation assays were performed to determine the effect of LMWHs on fibrillogenesis. RESULTS: Injection of mice with clinically relevant doses of LMWHs (enoxaparin and dalteparin) demonstrated a reduction in AA amyloid deposition. These compounds were capable of arresting the progression of AA amyloid and eventually resulting in regression of the amyloid deposits. In vitro analysis indicated that LMWHs prevented AA and Abeta peptide fibril formation by impeding the structural changes necessary for fibril formation. CONCLUSIONS: Our findings suggest that the LMWHs may provide beneficial effects against the development of amyloidoses, including Alzheimer's disease. AD - Department of Biochemistry, University of Kentucky, Lexington, Kentucky 40536, USA. FAU - Zhu, H AU - Zhu H FAU - Yu, J AU - Yu J FAU - Kindy, M S AU - Kindy MS LA - eng ID - AG12891/AG/NIA ID - NS31220/NS/NINDS PT - Journal Article CY - United States TA - Mol Med JID - 9501023 RN - 0 (Amyloid) RN - 0 (Enoxaparin) RN - 0 (Fibrinolytic Agents) RN - 0 (Tedelparin) SB - IM MH - Amyloid/*metabolism MH - Amyloidosis/*drug therapy/pathology MH - Animal MH - Circular Dichroism MH - Disease Models, Animal MH - Enoxaparin/*therapeutic use MH - Fibrinolytic Agents/*therapeutic use MH - Human MH - Mice MH - Mice, Inbred C57BL MH - Spleen/drug effects/pathology MH - Splenic Diseases/drug therapy/pathology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Tedelparin/*therapeutic use EDAT- 2001/10/10 10:00 MHDA- 2002/02/15 10:01 AID - S1528365801805176 [pii] PST - ppublish SO - Mol Med 2001 Aug;7(8):517-22. UI - 21486464 PMID- 11500500 DA - 20011015 DCOM- 20011204 LR - 20030103 IS - 0021-9258 VI - 276 IP - 42 DP - 2001 Oct 19 TI - New insights into the heparan sulfate proteoglycan-binding activity of apolipoprotein E. PG - 39138-44 AB - Defective binding of apolipoprotein E (apoE) to heparan sulfate proteoglycans (HSPGs) is associated with increased risk of atherosclerosis due to inefficient clearance of lipoprotein remnants by the liver. The interaction of apoE with HSPGs has also been implicated in the pathogenesis of Alzheimer's disease and may play a role in neuronal repair. To identify which residues in the heparin-binding site of apoE and which structural elements of heparan sulfate interact, we used a variety of approaches, including glycosaminoglycan specificity assays, (13)C nuclear magnetic resonance, and heparin affinity chromatography. The formation of the high affinity complex required Arg-142, Lys-143, Arg-145, Lys-146, and Arg-147 from apoE and N- and 6-O-sulfo groups of the glucosamine units from the heparin fragment. As shown by molecular modeling, using a high affinity binding octasaccharide fragment of heparin, these findings are consistent with a binding mode in which five saccharide residues of fully sulfated heparan sulfate lie in a shallow groove of the alpha-helix that contains the HSPG-binding site (helix 4 of the four-helix bundle of the 22-kDa fragment). This groove is lined with residues Arg-136, Ser-139, His-140, Arg-142, Lys-143, Arg-145, Lys-146, and Arg-147. In the model, all of these residues make direct contact with either the 2-O-sulfo groups of the iduronic acid monosaccharides or the N- and 6-O-sulfo groups of the glucosamine sulfate monosaccharides. This model indicates that apoE has an HSPG-binding site highly complementary to heparan sulfate rich in N- and O-sulfo groups such as that found in the liver and the brain. AD - Gladstone Institute of Cardiovascular Disease, San Francisco, California 94141, USA. FAU - Libeu, C P AU - Libeu CP FAU - Lund-Katz, S AU - Lund-Katz S FAU - Phillips, M C AU - Phillips MC FAU - Wehrli, S AU - Wehrli S FAU - Hernaiz, M J AU - Hernaiz MJ FAU - Capila, I AU - Capila I FAU - Linhardt, R J AU - Linhardt RJ FAU - Raffai, R L AU - Raffai RL FAU - Newhouse, Y M AU - Newhouse YM FAU - Zhou, F AU - Zhou F FAU - Weisgraber, K H AU - Weisgraber KH LA - eng ID - GM38060/GM/NIGMS ID - HL41633/HL/NHLBI ID - HL52622/HL/NHLBI ID - HL56083/HL/NHLBI PT - Journal Article CY - United States TA - J Biol Chem JID - 2985121R RN - 0 (Apolipoproteins E) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Polysaccharides) RN - 3416-24-8 (Glucosamine) RN - 56-45-1 (Serine) RN - 56-87-1 (Lysine) RN - 74-79-3 (Arginine) RN - 9005-49-6 (Heparin) RN - 9013-20-1 (Streptavidin) SB - IM MH - Animal MH - Apolipoproteins E/chemistry/*metabolism MH - Arginine/chemistry MH - Binding Sites MH - Biotinylation MH - Brain/metabolism MH - Cattle MH - Chromatography, Affinity MH - Dose-Response Relationship, Drug MH - Glucosamine/chemistry MH - Heparan Sulfate Proteoglycan/chemistry/*metabolism MH - Heparin/chemistry/metabolism MH - Human MH - Hydrogen-Ion Concentration MH - Kinetics MH - Liver/metabolism MH - Lysine/chemistry MH - Magnetic Resonance Spectroscopy MH - Models, Molecular MH - Mutation MH - Polysaccharides/metabolism MH - Protein Binding MH - Serine/chemistry MH - Streptavidin/chemistry MH - Support, U.S. Gov't, P.H.S. MH - Surface Plasmon Resonance MH - Time Factors EDAT- 2001/08/14 10:00 MHDA- 2002/01/05 10:01 PHST- 2001/Aug/10 [aheadofprint] AID - 10.1074/jbc.M104746200 [doi] AID - M104746200 [pii] PST - ppublish SO - J Biol Chem 2001 Oct 19;276(42):39138-44. UI - 21331285 PMID- 11437366 DA - 20010704 DCOM- 20010802 LR - 20011119 IS - 0006-291X VI - 285 IP - 1 DP - 2001 Jul 6 TI - Role of cysteine-291 and cysteine-322 in the polymerization of human tau into Alzheimer-like filaments. PG - 20-6 AB - Filamentous tau pathology is central to a large number of dementing disorders, including Alzheimer's disease in which polymerized tau is hyperphosphorylated. Previous studies on heparin-dependent tau polymerization, using recombinant tau isoforms lacking Cys-291, suggest that tau dimerization via Cys-322 is critical for initiation of assembly of soluble tau into filaments. We report heparin-dependent in vitro polymerization of human recombinant tau (1-383 isoform), containing both Cys-291 and Cys-322, into paired helical filaments as characterized by electron microscopy. Tau polymerization, under physiological tau concentrations in the presence of dithiothreitol (DTT), was followed by a Thioflavine S fluorescence assay. To understand the molecular basis for heparin-induced tau polymerization, we expressed and purified C291A, C322A, and C291A/C322A tau mutants. The DTT requirement for tau polymerization was abolished using either the C291A or C322A tau mutant and polymerization was not observed with the C291A/C322A tau double mutant. Analysis by sodium dodecyl sulfate gel electrophoresis showed that, unlike wild type tau, a significant amount of the C291A mutant and the C322A mutant is present as a disulfide bonded dimer. Taken together these results suggest that, in isoforms containing both Cys-291 and Cys-322, a dimeric tau with an intermolecular disulfide bond through either Cys-291 or Cys-322 is presumably acting as a seed for initiation of tau polymerization. CI - Copyright 2001 Academic Press. AD - Pharmacia Corporation, Kalamazoo, Michigan 49007, USA. FAU - Bhattacharya, K AU - Bhattacharya K FAU - Rank, K B AU - Rank KB FAU - Evans, D B AU - Evans DB FAU - Sharma, S K AU - Sharma SK LA - eng PT - Journal Article CY - United States TA - Biochem Biophys Res Commun JID - 0372516 RN - 0 (Biopolymers) RN - 0 (Protein Isoforms) RN - 0 (tau Proteins) RN - 3483-12-3 (Dithiothreitol) RN - 52-90-4 (Cysteine) SB - IM MH - Alzheimer Disease/*metabolism MH - Biopolymers/*metabolism MH - Cysteine/*metabolism MH - Dithiothreitol/chemistry MH - Electrophoresis, Polyacrylamide Gel MH - Human MH - Microscopy, Electron MH - Protein Isoforms/chemistry/*metabolism/ultrastructure MH - tau Proteins/chemistry/*metabolism/ultrastructure EDAT- 2001/07/05 10:00 MHDA- 2001/08/03 10:01 AID - 10.1006/bbrc.2001.5116 [doi] AID - S0006291X01951162 [pii] PST - ppublish SO - Biochem Biophys Res Commun 2001 Jul 6;285(1):20-6. UI - 21402949 PMID- 11423547 DA - 20010820 DCOM- 20010920 LR - 20030103 IS - 0021-9258 VI - 276 IP - 34 DP - 2001 Aug 24 TI - New insights on how metals disrupt amyloid beta-aggregation and their effects on amyloid-beta cytotoxicity. PG - 32293-9 AB - Amyloid-beta protein (A beta) aggregates in the brain to form senile plaques. By using thioflavin T, a dye that specifically binds to fibrillar structures, we found that metals such as Zn(II) and Cu(II) normally inhibit amyloid beta-aggregation. Another method for detecting A beta, which does not distinguish the types of aggregates, showed that these metals induce a non-beta-sheeted aggregation, as reported previously. Secondary structural analysis and microscopic studies revealed that metals induced A beta to make non-fibrillar aggregates by disrupting beta-sheet formation. These non-fibrillar A beta aggregates displayed much weaker Congo Red birefringence, and in separate cell culture experiments, were less toxic than self beta-aggregates, as demonstrated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The toxicity of soluble A beta was enhanced in the presence of Cu(II), which suggests the previously hypothesized role of A beta in generating oxidative stress. Finally, under an acidic condition, similar to that in the inflammation associated with senile plaques, beta-aggregation was robustly facilitated at one specific concentration of Zn(II) in the presence of heparin. However, because a higher concentration of Zn(II) virtually abolished this abnormal phenomenon, and at normal pH any concentrations strongly inhibit beta-aggregation and its associated cytotoxicity, including its anti-oxidative nature we suggest that Zn(II) has an overall protective effect against beta-amyloid toxicity. AD - Laboratory for Alzheimer's Disease, Brain Science Institute, RIKEN, Wako, Saitama 351-0198, Japan. FAU - Yoshiike, Y AU - Yoshiike Y FAU - Tanemura, K AU - Tanemura K FAU - Murayama, O AU - Murayama O FAU - Akagi, T AU - Akagi T FAU - Murayama, M AU - Murayama M FAU - Sato, S AU - Sato S FAU - Sun, X AU - Sun X FAU - Tanaka, N AU - Tanaka N FAU - Takashima, A AU - Takashima A LA - eng PT - Journal Article CY - United States TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid beta-Protein) RN - 7440-50-8 (Copper) RN - 7440-66-6 (Zinc) SB - IM MH - Amyloid beta-Protein/*metabolism/physiology MH - Animal MH - Cell Death/drug effects MH - Cell Line MH - Cell Survival/drug effects/*physiology MH - Cells, Cultured MH - Circular Dichroism MH - Copper/pharmacology MH - Hippocampus/cytology/drug effects MH - Human MH - Neurons/cytology/drug effects MH - Rats MH - Zinc/pharmacology EDAT- 2001/06/26 10:00 MHDA- 2001/09/21 10:01 PHST- 2001/Jun/22 [aheadofprint] AID - 10.1074/jbc.M010706200 [doi] AID - M010706200 [pii] PST - ppublish SO - J Biol Chem 2001 Aug 24;276(34):32293-9. UI - 21250702 PMID- 11352733 DA - 20010515 DCOM- 20010816 IS - 0006-2960 VI - 40 IP - 20 DP - 2001 May 22 TI - In vitro assembly of tau protein: mapping the regions involved in filament formation. PG - 5983-91 AB - Unraveling the mechanism of self-assembly of the protein tau into paired helical filaments (PHFs) is a crucial step toward the understanding of Alzheimer's and other neuropathological diseases at the molecular level. In an effort to map the role of different regions of tau in the mechanism of self-assembly, we have studied the polymerization ability of different tau fragments using an in vitro assay. Our results indicate that the N-terminal domain interferes with tau's ability to polymerize in vitro. The effect seems to be size dependent. Particularly, an isoform of tau from the peripheral nervous system, which has a much larger N-terminal domain, was found unable to form filaments in our in vitro assay. This finding can explain why in Alzheimer's patients PHFs only accumulate in the neurons from the central nervous system. We also report that a short segment of tau located in the third microtubule binding repeat (residues 317 to 335, peptide 1/2R) is probably the minimal segment of that region able to grow into filaments in vitro and in the presence of heparin. In contrast with whole peptide 1/2R, peptides corresponding to either the N-terminal or C-terminal halves of this segment were unable to form filaments. Finally, our polymerization studies of peptides from the C-terminal domain reveal a short sequence spanning residues 391 to 407 that grows into filaments in vitro. This tau segment forms filaments regardless of whether is incubated with heparin. Moreover, such filaments differ in diameter and morphology, suggesting a different mechanism of self-assembly. AD - Centro de Biologia Molecular "Severo Ochoa", Universidad Autonoma de Madrid, 28049-Madrid, Spain. FAU - Perez, M AU - Perez M FAU - Arrasate, M AU - Arrasate M FAU - Montejo De Garcini, E AU - Montejo De Garcini E FAU - Munoz, V AU - Munoz V FAU - Avila, J AU - Avila J LA - eng PT - Journal Article CY - United States TA - Biochemistry JID - 0370623 RN - 0 (Peptide Fragments) RN - 0 (Polymers) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Amino Acid Sequence MH - Amino Acid Substitution MH - Animal MH - Heparin/chemistry MH - Microfilaments/*chemistry/*metabolism/ultrastructure MH - Microscopy, Electron MH - Molecular Sequence Data MH - Peptide Fragments/chemistry/metabolism/ultrastructure MH - *Peptide Mapping/methods MH - Polymers/chemistry/metabolism MH - Rats MH - Repetitive Sequences, Amino Acid MH - Support, Non-U.S. Gov't MH - tau Proteins/*chemistry/*metabolism/ultrastructure EDAT- 2001/05/16 10:00 MHDA- 2001/08/17 10:01 AID - bi002961w [pii] PST - ppublish SO - Biochemistry 2001 May 22;40(20):5983-91. UI - 21159075 PMID- 11258893 DA - 20010322 DCOM- 20010510 IS - 0006-2960 VI - 40 IP - 9 DP - 2001 Mar 6 TI - Interaction of the N-terminal domain of apolipoprotein E4 with heparin. PG - 2826-34 AB - Apolipoprotein E (apoE) is an important lipid-transport protein in human plasma and brain. It has three common isoforms (apoE2, apoE3, and apoE4). ApoE is a major genetic risk factor in heart disease and in neurodegenerative disease, including Alzheimer's disease. The interaction of apoE with heparan sulfate proteoglycans plays an important role in lipoprotein remnant uptake and likely in atherogenesis and Alzheimer's disease. Here we report our studies of the interaction of the N-terminal domain of apoE4 (residues 1-191), which contains the major heparin-binding site, with an enzymatically prepared heparin oligosaccharide. Identified by its high affinity for the N-terminal domain of apoE4, this oligosaccharide was determined to be an octasaccharide of the structure DeltaUAp2S(1-->[4)-alpha-D-GlcNpS6S(1-->4)-alpha-L-IdoAp2S(1-->](3)4)-alph a-D-GlcNpS6S by nuclear magnetic resonance spectroscopy, capillary electrophoresis, and polyacrylamide gel electrophoresis. Kinetic analysis of the interaction between the N-terminal apoE4 fragment and immobilized heparin by surface plasmon resonance yielded a K(d) of 150 nM. A similar binding constant (K(d) = 140 nM) was observed for the interaction between immobilized N-terminal apoE4 and the octasaccharide. Isothermal titration calorimetry revealed a K(d) of 75 nM for the interaction of the N-terminal apoE fragment and the octasaccharide with a binding stoichiometry of approximately 1:1. Using previous studies and molecular modeling, we propose a binding site for this octasaccharide in a basic residue-rich region of helix 4 of the N-terminal fragment. From the X-ray crystal structure of the N-terminal apoE4, we predicted that binding of the octasaccharide at this site would result in a change in intrinsic fluorescence. This prediction was confirmed experimentally by an observed increase in fluorescence intensity with octasaccharide binding corresponding to a K(d) of approximately 1 microM. AD - Gladstone Institute of Cardiovascular Disease, Cardiovascular Research Institute, and Department of Pathology, University of California, San Francisco, California 94941-9100, USA. FAU - Dong, J AU - Dong J FAU - Peters-Libeu, C A AU - Peters-Libeu CA FAU - Weisgraber, K H AU - Weisgraber KH FAU - Segelke, B W AU - Segelke BW FAU - Rupp, B AU - Rupp B FAU - Capila, I AU - Capila I FAU - Hernaiz, M J AU - Hernaiz MJ FAU - LeBrun, L A AU - LeBrun LA FAU - Linhardt, R J AU - Linhardt RJ LA - eng SI - PDB/1B68 ID - GM 38060/GM/NIGMS ID - HL 41633/HL/NHLBI ID - HL 52622/HL/NHLBI ID - NS 35939/NS/NINDS PT - Journal Article CY - United States TA - Biochemistry JID - 0370623 RN - 0 (Apolipoproteins E) RN - 0 (Oligosaccharides) RN - 0 (Peptide Fragments) RN - 0 (apolipoprotein E-4) RN - 9005-49-6 (Heparin) SB - IM MH - Animal MH - Apolipoproteins E/chemistry/*metabolism MH - Calorimetry MH - Carbohydrate Sequence MH - Crystallography, X-Ray MH - Heparin/chemistry/*metabolism MH - Kinetics MH - Molecular Sequence Data MH - Oligosaccharides/chemistry/metabolism MH - Peptide Fragments/chemistry/*metabolism MH - Spectrometry, Fluorescence MH - Support, U.S. Gov't, Non-P.H.S. MH - Support, U.S. Gov't, P.H.S. MH - Surface Plasmon Resonance MH - Swine EDAT- 2001/03/22 10:00 MHDA- 2001/05/22 10:01 AID - bi002417n [pii] PST - ppublish SO - Biochemistry 2001 Mar 6;40(9):2826-34. UI - 21125850 PMID- 11106653 DA - 20010306 DCOM- 20010405 LR - 20030103 IS - 0021-9258 VI - 276 IP - 9 DP - 2001 Mar 2 TI - Amyloid-beta interactions with chondroitin sulfate-derived monosaccharides and disaccharides. implications for drug development. PG - 6412-9 AB - In Alzheimer's disease, the major pathological features are diffuse and senile plaques that are primarily composed of the amyloid-beta (A beta) peptide. It has been proposed that proteoglycans and glycosaminoglycans (GAG) facilitate amyloid fibril formation and/or stabilize the plaque aggregates. To develop effective therapeutics based on A beta-GAG interactions, understanding the A beta binding motif on the GAG chain is imperative. Using electron microscopy, fluorescence spectroscopy, and competitive inhibition ELISAs, we have evaluated the ability of chondroitin sulfate-derived monosaccharides and disaccharides to induce the structural changes in A beta that are associated with GAG interactions. Our results demonstrate that the disaccharides GalNAc-4-sulfate(4S), Delta UA-GalNAc-6-sulfate(6S), and Delta UA-GalNAc-4,6-sulfate(4S,6S), the iduronic acid-2-sulfate analogues, and the monosaccharides d-GalNAc-4S, d-GalNAc-6S, and d-GalNAc-4S,6S, but not d-GalNAc, d-GlcNAc, or Delta UA-GalNAc, induce the fibrillar features of A beta-GAG interactions. The binding affinities of all chondroitin sulfate-derived saccharides mimic those of the intact GAG chains. The sulfated monosaccharides and disaccharides compete with the intact chondroitin sulfate and heparin GAGs for A beta binding, as illustrated by competitive inhibition ELISAs. Therefore, the development of therapeutics based on the model of A beta-chondroitin sulfate binding may lead to effective inhibitors of the GAG-induced amyloid formation that is observed in vitro. AD - Centre for Research in Neurodegenerative Diseases, University of Toronto, Toronto, Ontario M5S 3H2, Canada. FAU - Fraser, P E AU - Fraser PE FAU - Darabie, A A AU - Darabie AA FAU - McLaurin, J A AU - McLaurin JA LA - eng PT - Journal Article CY - United States TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid beta-Protein) RN - 0 (Disaccharides) RN - 0 (Glycosaminoglycans) RN - 0 (Monosaccharides) RN - 9007-28-7 (Chondroitin Sulfates) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Amyloid beta-Protein/chemistry/*metabolism MH - Binding Sites MH - Chondroitin Sulfates/*metabolism MH - Disaccharides/*metabolism MH - Glycosaminoglycans/*metabolism MH - Heparitin Sulfate/metabolism MH - Human MH - Monosaccharides/*metabolism MH - Support, Non-U.S. Gov't EDAT- 2000/12/07 MHDA- 2001/04/06 10:01 PHST- 2000/Dec/05 [aheadofprint] AID - 10.1074/jbc.M008128200 [doi] AID - M008128200 [pii] PST - ppublish SO - J Biol Chem 2001 Mar 2;276(9):6412-9. UI - 20485509 PMID- 11029577 DA - 20001122 DCOM- 20001214 IS - 0014-2956 VI - 267 IP - 21 DP - 2000 Nov TI - Effect of amino-acid substitutions on Alzheimer's amyloid-beta peptide-glycosaminoglycan interactions. PG - 6353-61 AB - One of the major clinical features of Alzheimer's disease is the presence of extracellular amyloid plaques that are associated with glycosaminoglycan-containing proteoglycans. It has been proposed that proteoglycans and glycosaminoglycans facilitate amyloid fibril formation and/or stabilize these aggregates. Characterization of proteoglycan-protein interactions has suggested that basic amino acids in a specific conformation are necessary for glycosaminoglycan binding. Amyloid-beta peptide (Abeta) has a cluster of basic amino acids at the N-terminus (residues 13-16, His-His-Gln-Lys), which are considered critical for glycosaminoglycan interactions. To understand the molecular recognition of glycosaminoglycans by Abeta, we have examined a series of synthetic peptides with systematic alanine substitutions. These include: His13-->Ala, His14-->Ala, Lys16-->Ala, His13His14Lys16-->Ala and Arg5His6-->Ala. Alanine substitutions result in differences in both the secondary and fibrous structure of Abeta1-28 as determined by circular dichroism spectroscopy and electron microscopy. The results demonstrate that the His-His-Gln-Lys region of Abeta, and in particular His13, is an important structural domain, as Ala substitution produces a dysfunctional folding mutant. Interaction of the substituted peptides with heparin and chondroitin sulfate glycosaminoglycans demonstrate that although electrostatic interactions contribute to binding, nonionic interactions such as hydrogen bonding and van der Waals packing play a role in glycosaminoglycan-induced Abeta folding and aggregation. AD - Centre for Research in Neurodegenerative Diseases, Department of Laboratory Medicine , University of Toronto, Toronto, Ontario, Canada. j.mclaurin@utoronto.ca FAU - McLaurin, J AU - McLaurin J FAU - Fraser, P E AU - Fraser PE LA - eng PT - Journal Article CY - GERMANY TA - Eur J Biochem JID - 0107600 RN - 0 (Amyloid beta-Protein) RN - 0 (Glycosaminoglycans) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta protein (1-28)) RN - 0 (amyloid beta-protein (1-40)) RN - 0 (beta-amyloid (1-42)) RN - 24967-94-0 (Dermatan Sulfate) RN - 56-41-7 (Alanine) RN - 9005-49-6 (Heparin) RN - 9007-28-7 (Chondroitin Sulfates) SB - IM MH - Alanine/genetics/metabolism MH - Alzheimer Disease/genetics/*metabolism MH - Amino Acid Sequence MH - Amino Acid Substitution/*genetics MH - Amyloid beta-Protein/*chemistry/genetics/*metabolism/ultrastructure MH - Chondroitin Sulfates/metabolism MH - Circular Dichroism MH - Dermatan Sulfate/metabolism MH - Electrostatics MH - Glycosaminoglycans/*metabolism/ultrastructure MH - Heparin/metabolism MH - Human MH - Hydrogen Bonding MH - Microscopy, Electron MH - Molecular Sequence Data MH - Mutation/genetics MH - Peptide Fragments/*chemistry/genetics/*metabolism/ultrastructure MH - Protein Binding MH - Protein Folding MH - Protein Structure, Quaternary MH - Protein Structure, Secondary MH - Support, Non-U.S. Gov't EDAT- 2000/10/13 11:00 MHDA- 2001/02/28 10:01 AID - ejb1725 [pii] PST - ppublish SO - Eur J Biochem 2000 Nov;267(21):6353-61. UI - 20545467 PMID- 10973954 DA - 20001218 DCOM- 20010111 IS - 0021-9258 VI - 275 IP - 48 DP - 2000 Dec 1 TI - SERPIN regulation of factor XIa. The novel observation that protease nexin 1 in the presence of heparin is a more potent inhibitor of factor XIa than C1 inhibitor. PG - 37340-6 AB - In the present studies we have made the novel observation that protease nexin 1 (PN1), a member of the serine protease inhibitor (SERPIN) superfamily, is a potent inhibitor of the blood coagulation Factor XIa (FXIa). The inhibitory complexes formed between PN1 and FXIa are stable when subjected to reducing agents, SDS, and boiling, a characteristic of the acyl linkage formed between SERPINs and their cognate proteases. Using a sensitive fluorescence-quenched peptide substrate, the K(assoc) of PN1 for FXIa was determined to be 7.9 x 10(4) m(-)(1) s(-)(1) in the absence of heparin. In the presence of heparin, this rate was accelerated to 1.7 x 10(6), M(-)(1) s(-)(1), making PN1 a far better inhibitor of FXIa than C1 inhibitor, which is the only other SERPIN known to significantly inhibit FXIa. FXIa-PN1 complexes are shown to be internalized and degraded by human fibroblasts, most likely via the low density lipoprotein receptor-related protein (LRP), since degradation was strongly inhibited by the LRP agonist, receptor-associated protein. Since FXIa proteolytically modifies the amyloid precursor protein, this observation may suggest an accessory role for PN1 in the pathobiogenesis of Alzheimer's disease. AD - Department of Developmental and Cell Biology, School of Biological Sciences, University of California, Irvine, California 92627, USA. FAU - Knauer, D J AU - Knauer DJ FAU - Majumdar, D AU - Majumdar D FAU - Fong, P C AU - Fong PC FAU - Knauer, M F AU - Knauer MF LA - eng ID - RO1GM34001-12/GM/NIGMS PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (C1-inhibitor protein) RN - 0 (Carrier Proteins) RN - 0 (Complement 1 Inactivators) RN - 0 (Serine Proteinase Inhibitors) RN - 0 (Serpins) RN - 0 (protease-nexin) RN - 9005-49-6 (Heparin) RN - EC 3.4.21.27 (Factor XIa) SB - IM MH - Carrier Proteins/*pharmacology MH - Cell Line MH - Complement 1 Inactivators/*pharmacology MH - Factor XIa/antagonists & inhibitors/*physiology MH - Heparin/*pharmacology MH - Human MH - Serine Proteinase Inhibitors/pharmacology MH - Serpins/*physiology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 2000/09/07 11:00 MHDA- 2001/02/28 10:01 AID - 10.1074/jbc.M003909200 [doi] AID - M003909200 [pii] PST - ppublish SO - J Biol Chem 2000 Dec 1;275(48):37340-6. UI - 20437008 PMID- 10899436 DA - 20000925 DCOM- 20000925 LR - 20001218 IS - 0006-3002 VI - 1502 IP - 1 DP - 2000 Jul 26 TI - Tau mutations in frontotemporal dementia FTDP-17 and their relevance for Alzheimer's disease. PG - 110-21 AB - Alzheimer's disease is characterised by the degeneration of selected populations of nerve cells that develop filamentous inclusions prior to degeneration. The neuronal inclusions of Alzheimer's disease are made of the microtubule-associated protein tau, in a hyperphosphorylated state. Abundant filamentous tau inclusions are not limited to Alzheimer's disease. They are the defining neuropathological characteristic of frontotemporal dementias, such as Pick's disease, and of progressive supranuclear palsy and corticobasal degeneration. The discovery of mutations in the tau gene in familial frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) has provided a direct link between tau dysfunction and dementing disease. Known mutations produce either a reduced ability of tau to interact with microtubules, or an overproduction of tau isoforms with four microtubule-binding repeats. This leads in turn to the assembly of tau into filaments similar or identical to those found in Alzheimer's disease brain. Several missense mutations also have a stimulatory effect on heparin-induced tau filament formation. Assembly of tau into filaments may be the gain of toxic function that is believed to underlie the demise of affected brain cells. AD - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK. mg@mrc-lmb.cam.ac.uk FAU - Goedert, M AU - Goedert M FAU - Spillantini, M G AU - Spillantini MG LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - NETHERLANDS TA - Biochim Biophys Acta JID - 0217513 RN - 0 (FTDP-17 protein) RN - 0 (Microtubule-Associated Proteins) RN - 0 (Phosphoproteins) RN - 0 (Protein Isoforms) RN - 0 (tau Proteins) SB - IM MH - Age of Onset MH - Alzheimer Disease/*genetics/pathology MH - Brain/metabolism/pathology MH - Dementia/*genetics/pathology MH - Exons MH - Human MH - Introns MH - Microtubule-Associated Proteins/*genetics MH - Microtubules/ultrastructure MH - *Mutation MH - Phosphoproteins/metabolism MH - Protein Isoforms/metabolism MH - tau Proteins/*genetics/metabolism/ultrastructure RF - 103 EDAT- 2000/07/19 11:00 MHDA- 2000/09/30 11:01 AID - S0925443900000375 [pii] PST - ppublish SO - Biochim Biophys Acta 2000 Jul 26;1502(1):110-21. UI - 20351567 PMID- 10891598 DA - 20000925 DCOM- 20000925 LR - 20021213 IS - 0169-328X VI - 78 IP - 1-2 DP - 2000 May 31 TI - Binding specificity of monoclonal antibody AD2: influence of the phosphorylation state of tau. PG - 181-5 AB - Using recombinant human tau protein phosphorylated by a brain extract and the glycogen synthase kinase-3beta in the absence or the presence of heparin, we showed that phosphorylation-dependent antibody AD2 recognition only requires phosphorylated Ser-396. By the Spot multiple peptide synthesis method, we showed that Tyr-394, Ser(P)-396 and Pro-397 are critical for AD2 binding. A decrease in the binding of AD2 was observed with increasing phosphorylation of residues in the vicinity of Ser(P)-396. AD - CNRS UMR 5094 Institut de Biotechnologie en Immunoanalyse et Pharmacologie UFR Pharmacie, 15 Av. Charles Flahault, 34060 Montpellier Cedex 2, France. FAU - Torreilles, F AU - Torreilles F FAU - Roquet, F AU - Roquet F FAU - Granier, C AU - Granier C FAU - Pau, B AU - Pau B FAU - Mourton-Gilles, C AU - Mourton-Gilles C LA - eng PT - Journal Article CY - NETHERLANDS TA - Brain Res Mol Brain Res JID - 8908640 RN - 0 (Antibodies, Monoclonal) RN - 0 (Anticoagulants) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) SB - IM MH - Alzheimer Disease/metabolism MH - Amino Acid Sequence MH - Antibodies, Monoclonal/*immunology MH - *Antibody Specificity MH - Anticoagulants/pharmacology MH - Brain Chemistry MH - Ca(2+)-Calmodulin Dependent Protein Kinase/metabolism MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Heparin/pharmacology MH - Human MH - Neurons/chemistry/enzymology MH - Phosphorylation MH - Protein Binding/drug effects/immunology MH - tau Proteins/chemistry/*immunology/*metabolism EDAT- 2000/07/13 11:00 MHDA- 2000/09/30 11:01 AID - S0169328X00000735 [pii] PST - ppublish SO - Brain Res Mol Brain Res 2000 May 31;78(1-2):181-5. UI - 20378979 PMID- 10806211 DA - 20000907 DCOM- 20000907 LR - 20020419 IS - 0021-9258 VI - 275 IP - 31 DP - 2000 Aug 4 TI - Phosphorylation of the beta-amyloid precursor protein at the cell surface by ectocasein kinases 1 and 2. PG - 23523-9 AB - The beta-amyloid precursor protein (betaAPP) is one of the rare proteins known to be phosphorylated within its ectodomain. We have shown previously that betaAPP can be phosphorylated within secretory vesicles and at the cell surface (Walter, J., Capell, A., Hung, A. Y. , Langen, H., Schnolzer, M., Thinakaran, G., Sisodia, S. S., Selkoe, D. J., and Haass, C. (1997) J. Biol. Chem. 272, 1896-1903). We have now specifically characterized the phosphorylation of cell surface-located betaAPP and identified two ectoprotein kinases that phosphorylate betaAPP at the outer face of the plasma membrane. By using selective protein kinase inhibitors and by investigating the usage of ATP and GTP as cosubstrates, we demonstrate that membrane-bound betaAPP as well as secreted forms of betaAPP can be phosphorylated by casein kinase (CK) 1- and CK2-like ectoprotein kinases. The ectodomain of betaAPP was also phosphorylated by purified CK1 and CK2 in vitro, but not by protein kinases A and C. Phosphorylation of betaAPP by ectoprotein kinases and by purified CK1 and CK2 occurred within an acidic domain in the N-terminal half of the protein. Heparin strongly inhibited the phosphorylation of cell-surface betaAPP by ecto-CK1 and ecto-CK2, indicating a regulatory role of this extracellular matrix component in betaAPP phosphorylation. AD - Adolf-Butenandt-Institut, Department of Biochemistry, Laboratory for Alzheimer's Disease Research, Ludwig-Maximilians-Universat Munchen, Schillerstrasse 44, D-80336 Munich, Germany. jwalter@pbm.med.uni-muenchen.de FAU - Walter, J AU - Walter J FAU - Schindzielorz, A AU - Schindzielorz A FAU - Hartung, B AU - Hartung B FAU - Haass, C AU - Haass C LA - eng PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid beta-Protein Precursor) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.- (casein kinase) RN - EC 2.7.1.- (casein kinase II) RN - EC 2.7.1.- (ectoprotein kinase) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 2.7.1.37 (Protein-Serine-Threonine Kinases) SB - IM MH - Amyloid beta-Protein Precursor/*metabolism MH - Cell Membrane/*metabolism MH - Heparin/pharmacology MH - Human MH - Models, Biological MH - Phosphorylation/drug effects MH - Protein Kinases/*metabolism MH - Protein Structure, Tertiary MH - Protein-Serine-Threonine Kinases/*metabolism MH - Support, Non-U.S. Gov't EDAT- 2000/05/12 09:00 MHDA- 2000/09/09 11:01 AID - 10.1074/jbc.M002850200 [doi] AID - M002850200 [pii] PST - ppublish SO - J Biol Chem 2000 Aug 4;275(31):23523-9. UI - 20229636 PMID- 10765057 DA - 20000606 DCOM- 20000606 LR - 20021101 IS - 0014-3022 VI - 43 IP - 3 DP - 2000 TI - Age-dependent decline in the apolipoprotein E level in cerebrospinal fluid from control subjects and its increase in cerebrospinal fluid from patients with Alzheimer's disease. PG - 161-9 AB - In order to address the significance of apolipoprotein E (apoE) in the pathogenesis as well as the clinical diagnosis of Alzheimer's disease (AD), we measured its level in cerebrospinal fluid (CSF) from randomly selected Japanese control subjects at various ages (n = 36), which included 14 age-matched controls, and from AD patients including early-onset (n = 11, EOAD) and late-onset (n = 14, LOAD) cases. The CSF apoE level in controls linearly decreased during aging to over 80 years (r(2) = 0.323, p < 0.0001). The CSF apoE level in AD patients was 31.9% elevated compared to the age-matched controls (n = 14, p < 0.05) and linearly increased with a decrement of the patients' Mini Mental State Examination scores. Moreover, the CSF apoE level of EOAD patients (n = 11) was higher than that of LOAD patients (n = 14, p < 0.05), whose APOE epsilon4 allele frequency was significantly higher than that of controls (chi(2) = 7. 16, p < 0.03). Two-dimensional gel electrophoretic analysis of the heparin-Mn(2+)-precipitable lipoprotein fraction in CSFs showed that the ratio between the level of CSF apoA-I and that of CSF apoE of controls was significantly higher than those of all AD and LOAD subjects (p < 0.01, p < 0.05), while the CSF apoA-I-to-apoE ratios of the two AD groups were not significantly different. These results suggest that overproduction of apoE protein may be a consequence of astroglial response to neurodegeneration in AD and that the determination of CSF apoliprotein levels serves as a clinical marker for monitoring the progression of AD. CI - Copyright 2000 S. Karger AG, Basel AD - Department of Dynamic Pathology, Research Institute for Neurological Diseases and Geriatrics, Kyoto Prefectural University of Medicine, Kyoto, Japan. FAU - Fukuyama, R AU - Fukuyama R FAU - Mizuno, T AU - Mizuno T FAU - Mori, S AU - Mori S FAU - Yanagisawa, K AU - Yanagisawa K FAU - Nakajima, K AU - Nakajima K FAU - Fushiki, S AU - Fushiki S LA - eng PT - Journal Article CY - SWITZERLAND TA - Eur Neurol JID - 0150760 RN - 0 (Apolipoproteins E) SB - IM MH - Adolescent MH - Adult MH - Age Factors MH - Aged MH - Aged, 80 and over MH - Alzheimer Disease/cerebrospinal fluid/*diagnosis/genetics MH - Apolipoproteins E/*cerebrospinal fluid/genetics MH - Child MH - Electrophoresis, Gel, Two-Dimensional MH - Enzyme-Linked Immunosorbent Assay MH - Female MH - Genotype MH - Human MH - Male MH - Middle Age MH - Reference Values EDAT- 2000/04/15 09:00 MHDA- 2000/06/10 09:00 AID - ene43161 [pii] PST - ppublish SO - Eur Neurol 2000;43(3):161-9. UI - 20199877 PMID- 10737616 DA - 20000411 DCOM- 20000411 LR - 20021213 IS - 0022-3042 VI - 74 IP - 4 DP - 2000 Apr TI - Phosphorylation sites on tau identified by nanoelectrospray mass spectrometry: differences in vitro between the mitogen-activated protein kinases ERK2, c-Jun N-terminal kinase and P38, and glycogen synthase kinase-3beta. PG - 1587-95 AB - The stress-activated kinases c-Jun N-terminal kinase (JNK) and p38 are members of the mitogen-activated protein (MAP) kinase family and take part in signalling cascades initiated by various forms of stress. Their targets include the microtubule-associated protein tau, which becomes hyperphosphorylated in Alzheimer's disease. It is necessary, as a forerunner for in vivo studies, to identify the protein kinases and phosphatases that are responsible for phosphate turnover at individual sites. Using nanoelectrospray mass spectrometry, we have undertaken an extensive comparison of phosphorylation in vitro by several candidate tau kinases, namely, JNK, p38, ERK2, and glycogen synthase kinase 3beta (GSK3beta). Between 10 and 15 sites were identified for each kinase. The three MAP kinases phosphorylated Ser202 and Thr205 but not detectably Ser199, whereas conversely GSK3beta phosphorylated Ser199 but not detectably Ser202 or Thr205. Phosphorylated Ser404 was found with all of these kinases except JNK. The MAP kinases may not be strictly proline specific: p38 phosphorylated the nonproline sites Ser185, Thr245, Ser305, and Ser356, whereas ERK2 was the most strict. All of the sites detected except Thr245 and Ser305 are known or suspected phosphorylation sites in paired helical filament-tau extracted from Alzheimer brains. Thus, the three MAP kinases and GSK3beta are importantly all strong candidates as tau kinases that may be involved in the pathogenic hyperphosphorylation of tau in Alzheimer's disease. AD - Department of Neuroscience, Institute of Psychiatry, King's College London, England. h.reynolds@iop.kcl.ac.uk FAU - Reynolds, C H AU - Reynolds CH FAU - Betts, J C AU - Betts JC FAU - Blackstock, W P AU - Blackstock WP FAU - Nebreda, A R AU - Nebreda AR FAU - Anderton, B H AU - Anderton BH LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Anticoagulants) RN - 0 (tau Proteins) RN - 147-85-3 (Proline) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) RN - EC 2.7.1.37 (Mitogen-Activated Protein Kinases) RN - EC 2.7.1.37 (p42 MAP Kinase) RN - EC 2.7.10.- (c-Jun amino-terminal kinase) RN - EC 2.7.10.- (mitogen-activated protein kinase p38) SB - IM MH - Alzheimer Disease/metabolism MH - Amino Acid Sequence MH - Animal MH - Anticoagulants/pharmacology MH - Binding Sites/drug effects/physiology MH - Ca(2+)-Calmodulin Dependent Protein Kinase/*metabolism MH - Cells, Cultured MH - Comparative Study MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Heparin/pharmacology MH - In Vitro MH - Insects MH - Mitogen-Activated Protein Kinases/*metabolism MH - Molecular Sequence Data MH - Neurofibrillary Tangles/enzymology MH - Phosphorylation MH - Proline MH - Signal Transduction/physiology MH - Spectrum Analysis, Mass MH - Support, Non-U.S. Gov't MH - p42 MAP Kinase/metabolism MH - tau Proteins/chemistry/*metabolism EDAT- 2000/03/29 09:00 MHDA- 2000/04/15 09:00 PST - ppublish SO - J Neurochem 2000 Apr;74(4):1587-95. UI - 20096612 PMID- 10629403 DA - 20000225 DCOM- 20000225 LR - 20021029 IS - 0301-0147 VI - 29 Suppl S1 DP - 1999 Dec TI - What else can 'Heparin' do? PG - 38-47 AB - This review article begins by discussing the molecular basis of the blood anticoagulant effect of heparin and some species of heparan sulphate (HS). A highly specific pentasaccharide sequence, containing a glucosamine 3-O-sulphate group, is a key structural element for this action. The biosynthesis of heparin and HS is outlined. Different types of HS proteoglycans exist. Analysis of HS preparations from different mammalian organs has indicated that the structural variability of the polysaccharide is due to regulated polymer modification. In addition to antithrombin, HS chains bind a very large number of other proteins in vivo. Such binding often appears to depend on the presence of specific sequences of different monosaccharide building-blocks and has diverse implications. Many physiological and pathological processes in the mammalian body appear to be influenced or regulated by HS proteoglycans. For example, the proper assembly of HS chains is believed to play an important role in normal embryonic and mammalian development. Diseases such as diabetes, amyloidosis and Alzheimer's may be associated with changes in HS structure. Finally, the possibilities and strategies for developing drugs based on HS chemistry are discussed. CI - Copyright 1999 S. Karger AG, Basel AD - Department of Medical Biochemistry and Microbiology, The Biomedical Centre, University of Uppsala, Uppsala, Sweden. FAU - Lindahl, U AU - Lindahl U LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - SWITZERLAND TA - Haemostasis JID - 0371574 RN - 0 (Anticoagulants) RN - 9005-49-6 (Heparin) SB - IM MH - Animal MH - Anticoagulants/chemistry/*pharmacology MH - Female MH - Heparin/chemistry/*pharmacology MH - Human MH - Pregnancy MH - Protein Binding MH - Structure-Activity Relationship RF - 44 EDAT- 2000/01/12 09:00 MHDA- 2000/03/04 09:00 AID - hae9a038 [pii] PST - ppublish SO - Haemostasis 1999 Dec;29 Suppl S1:38-47. UI - 20074206 PMID- 10608507 DA - 20000112 DCOM- 20000112 LR - 20001218 IS - 1065-6251 VI - 7 IP - 1 DP - 2000 Jan TI - Expression and function of serum amyloid A, a major acute-phase protein, in normal and disease states. PG - 64-9 AB - Serum amyloid A (SAA), the precursor protein in inflammation-associated reactive amyloidosis (AA-type), is an acute phase reactant whose level in the blood increases in response to various insults. It is expressed in the liver, but its physiological role is not well understood. Recently, a broader view of SAA expression and function has been emerging. Expression studies show local production of SAA proteins in histologically normal, atherosclerotic, Alzheimer, inflammatory, and tumor tissues. Binding sites in the SAA protein for high density lipoproteins, calcium, laminin, and heparin/heparan-sulfate were described. Adhesion motifs were identified and new functions, affecting cell adhesion, migration, proliferation and aggregation have been described. These findings emphasize the importance of SAA in various physiological and pathological processes, including inflammation, atherosclerosis, thrombosis, AA-amyloidosis, rheumatoid arthritis, and neoplasia. In addition, recent experiments suggest that SAA may play a "housekeeping" role in normal human tissues. AD - Hematology Unit, Hadassah University Hospital, Mount Scopus, Jerusalem, Israel. matzner@cc.huji.ac.il FAU - Urieli-Shoval, S AU - Urieli-Shoval S FAU - Linke, R P AU - Linke RP FAU - Matzner, Y AU - Matzner Y LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Curr Opin Hematol JID - 9430802 RN - 0 (Acute-Phase Proteins) RN - 0 (Amyloid Protein AA) RN - 0 (Apolipoproteins) RN - 0 (Protein Precursors) RN - 0 (amyloid protein AA precursor) SB - IM MH - Acute-Phase Proteins/*physiology MH - Amyloid Protein AA/chemistry/*genetics/*physiology MH - Apolipoproteins/chemistry/*genetics/*physiology MH - Human MH - Protein Precursors MH - Support, Non-U.S. Gov't RF - 71 EDAT- 1999/12/23 MHDA- 1999/12/23 00:01 PST - ppublish SO - Curr Opin Hematol 2000 Jan;7(1):64-9. UI - 20050625 PMID- 10583407 DA - 20000131 DCOM- 20000131 LR - 20001218 IS - 0014-2956 VI - 266 IP - 3 DP - 1999 Dec TI - Interactions of Alzheimer amyloid-beta peptides with glycosaminoglycans effects on fibril nucleation and growth. PG - 1101-10 AB - Proteoglycans and their constituent glycosaminoglycans are associated with all amyloid deposits and may be involved in the amyloidogenic pathway. In Alzheimer's disease, plaques are composed of the amyloid-beta peptide and are associated with at least four different proteoglycans. Using CD spectroscopy, fluorescence spectroscopy and electron microscopy, we examined glycosaminoglycan interaction with the amyloid-beta peptides 1-40 (Abeta40) and 1-42 (Abeta42) to determine the effects on peptide conformation and fibril formation. Monomeric amyloid-beta peptides in trifluoroethanol, when diluted in aqueous buffer, undergo a slow random to amyloidogenic beta sheet transition. In the presence of heparin, heparan sulfate, keratan sulfate or chondroitin sulfates, this transition was accelerated with Abeta42 rapidly adopting a beta-sheet conformation. This was accompanied by the appearance of well-defined amyloid fibrils indicating an enhanced nucleation of Abeta42. Incubation of preformed Abeta42 fibrils with glycosaminoglycans resulted in extensive lateral aggregation and precipitation of the fibrils. The glycosaminoglycans differed in their relative activities with the chondroitin sulfates producing the most pronounced effects. The less amyloidogenic Abeta40 isoform did not show an immediate structural transition that was dependent upon the shielding effect by the phosphate counter ion. Removal or substitution of phosphate resulted in similar glycosaminoglycan-induced conformational and aggregation changes. These findings clearly demonstrate that glycosaminoglycans act at the earliest stage of fibril formation, namely amyloid-beta nucleation, and are not simply involved in the lateral aggregation of preformed fibrils or nonspecific adhesion to plaques. The identification of a structure-activity relationship between amyloid-beta and the different glycosaminoglycans, as well as the condition dependence for glycosaminoglycan binding, are important for the successful development and evaluation of glycosaminoglycan-specific therapeutic interventions. AD - Centre for Research in Neurodegenerative Diseases, University of Toronto, Ontario, Canada. j.mclaurin@utoronto.ca FAU - McLaurin, J AU - McLaurin J FAU - Franklin, T AU - Franklin T FAU - Zhang, X AU - Zhang X FAU - Deng, J AU - Deng J FAU - Fraser, P E AU - Fraser PE LA - eng PT - Journal Article CY - GERMANY TA - Eur J Biochem JID - 0107600 RN - 0 (Amyloid beta-Protein) RN - 0 (Glycosaminoglycans) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta-protein (1-40)) RN - 0 (beta-amyloid (1-42)) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/*chemistry/*metabolism/ultrastructure MH - Circular Dichroism MH - Glycosaminoglycans/*chemistry/*metabolism MH - Human MH - Hydrogen-Ion Concentration MH - In Vitro MH - Microscopy, Electron MH - Molecular Sequence Data MH - Peptide Fragments/*chemistry/*metabolism/ultrastructure MH - Protein Conformation MH - Support, Non-U.S. Gov't EDAT- 1999/12/03 MHDA- 1999/12/03 00:01 AID - ejb957 [pii] PST - ppublish SO - Eur J Biochem 1999 Dec;266(3):1101-10. UI - 20002860 PMID- 10529469 DA - 19991206 DCOM- 19991206 LR - 20011102 IS - 0169-328X VI - 72 IP - 2 DP - 1999 Oct 1 TI - Phosphorylation of mitogen-activated protein kinase is altered in neuroectodermal cells overexpressing the human amyloid precursor protein 751 isoform. PG - 115-20 AB - The aberrant expression or processing of the amyloid precursor protein (APP) is the only known genetic basis for presenile familial Alzheimer's disease, and the molecular connection between APP and tau has been perplexing. Attention has focused on proline-directed serine/threonine kinases as mediating the cytoskeletal modifications of Alzheimer's disease, and we show that overexpression of APP can influence the activation of a candidate kinase, the mitogen-activated protein kinase (MAPK). In murine embryonal carcinoma cells stably transfected with the human 751 isoform of APP, we observed steady-state hyperactivation of p42(MAPK) concomitant with APP overexpression 3 days after neuroectodermal differentiation. In more mature differentiated cells, immunocytochemical analysis revealed enhanced basal somatic and nuclear immunoreactivity for phosphorylated MAPK coupled with an attenuated phosphorylation response to growth factor stimulation. Our results suggest that APP can influence the MAPK signaling pathway in such a way that the absolute and time-dependent activation required for discrimination of the appropriate downstream response are compromised. Such an effect would have important consequences for the functioning of cells coincidentally expressing both proteins, a situation that occurs in neuronal populations vulnerable to Alzheimer's disease pathology. AD - Department of Pharmacology and Therapeutics, McGill University, 3655 Drummond Street, Room 1325, Montreal, QC, Canada. FAU - Grant, S M AU - Grant SM FAU - Morinville, A AU - Morinville A FAU - Maysinger, D AU - Maysinger D FAU - Szyf, M AU - Szyf M FAU - Cuello, A C AU - Cuello AC LA - eng ID - AG-11093/AG/NIA PT - Journal Article CY - NETHERLANDS TA - Brain Res Mol Brain Res JID - 8908640 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (MAP Kinase Signaling System) RN - 0 (Protein Isoforms) RN - 0 (Recombinant Fusion Proteins) RN - 104781-85-3 (Fibroblast Growth Factor 1) RN - 302-79-4 (Tretinoin) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.37 (p42 MAP Kinase) SB - IM MH - Amyloid beta-Protein Precursor/genetics/*physiology MH - Animal MH - Carcinoma, Embryonal/pathology MH - Cell Differentiation/drug effects MH - Ectoderm/cytology/*drug effects MH - Fibroblast Growth Factor 1/pharmacology MH - Heparin/pharmacology MH - Human MH - MAP Kinase Signaling System/drug effects/*physiology MH - Mice MH - Phosphorylation MH - Protein Isoforms/genetics/*physiology MH - Protein Processing, Post-Translational MH - Recombinant Fusion Proteins/pharmacology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Transfection MH - Tretinoin/pharmacology MH - Tumor Cells, Cultured MH - p42 MAP Kinase/*physiology EDAT- 1999/10/26 MHDA- 1999/10/26 00:01 AID - S0169328X99001576 [pii] PST - ppublish SO - Brain Res Mol Brain Res 1999 Oct 1;72(2):115-20. UI - 99396227 PMID- 10468151 DA - 19991005 DCOM- 19991005 LR - 20001218 IS - 1065-6251 VI - 6 IP - 5 DP - 1999 Sep TI - Laboratory markers of platelet activation and their clinical significance. PG - 342-8 AB - Whole blood flow cytometry is a powerful new laboratory technique for assessment of platelet activation and function. Flow cytometry can be used to measure platelet hyperreactivity, circulating activated platelets, leukocyte-platelet aggregates, and procoagulant platelet-derived microparticles in a number of clinical settings, including acute coronary syndromes, angioplasty, cardiopulmonary bypass, acute cerebrovascular ischemia, peripheral vascular disease, diabetes mellitus, preeclampsia, and Alzheimer's disease. Clinical applications of whole blood flow cytometric assays of platelet function in these diseases may include identification of patients who would benefit from additional antiplatelet therapy and prediction of ischemic events. Circulating monocyte-platelet aggregates appear to be a more sensitive marker of in vivo platelet activation than circulating P-selectin-positive platelets. Flow cytometry can also be used in the following clinical settings: monitoring of glycoprotein IIb-IIIa antagonist therapy, diagnosis of inherited deficiencies of platelet surface glycoproteins, diagnosis of storage pool disease, diagnosis of heparin-induced thrombocytopenia, and measurement of the rate of thrombopoiesis. AD - Center for Platelet Function Studies, Department of Pediatrics, University of Massachusetts Medical School, Worcester 01655, USA. michelson@platelets.org FAU - Michelson, A D AU - Michelson AD FAU - Furman, M I AU - Furman MI LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Curr Opin Hematol JID - 9430802 RN - 0 (Biological Markers) RN - 0 (Platelet Aggregation Inhibitors) RN - 0 (Platelet Glycoprotein GPIIb-IIIa Complex) SB - IM MH - Biological Markers/*blood MH - Cerebrovascular Disorders/blood/drug therapy MH - Coronary Disease/blood/drug therapy MH - Flow Cytometry MH - Human MH - Platelet Activation/*drug effects MH - Platelet Aggregation Inhibitors/*therapeutic use MH - Platelet Glycoprotein GPIIb-IIIa Complex/*antagonists & inhibitors RF - 60 EDAT- 1999/09/01 MHDA- 1999/09/01 00:01 PST - ppublish SO - Curr Opin Hematol 1999 Sep;6(5):342-8. UI - 99362155 PMID- 10435777 DA - 19990920 DCOM- 19990920 LR - 20001218 IS - 0361-9230 VI - 49 IP - 3 DP - 1999 Jun TI - Inhibitors of intracellular phospholipase A2 activity: their neurochemical effects and therapeutical importance for neurological disorders. PG - 139-53 AB - Intracellular phospholipases A2 (PLA2) are a diverse group of enzymes with a growing number of members. These enzymes hydrolyze membrane phospholipids into fatty acid and lysophospholipids. These lipid products may serve as intracellular second messengers or can be further metabolized to potent inflammatory mediators, such as eicosanoids and platelet-activating factors. Several inhibitors of nonneural intracellular PLA2 have been recently discovered. However, nothing is known about their neurochemical effects, mechanism of action or toxicity in human or animal models of neurological disorders. Elevated intracellular PLA2 activities, found in neurological disorders strongly associated with inflammation and oxidative stress (ischemia, spinal cord injury, and Alzheimer's disease), can be treated with specific, potent and nontoxic inhibitors of PLA2 that can cross blood-brain barrier without harm. Currently, potent intracellular PLA2 inhibitors are not available for clinical use in human or animal models of neurological disorders, but studies on this interesting topic are beginning to emerge. The use of nonspecific intracellular PLA2 inhibitors (quinacrine, heparin, gangliosides, vitamin E) in animal model studies of neurological disorders in vivo has provided some useful information on tolerance, toxicity, and effectiveness of these compounds. AD - Department of Medical Biochemistry, The Ohio State University, Columbus 43210, USA. farooqui.l@osu.edu FAU - Farooqui, A A AU - Farooqui AA FAU - Litsky, M L AU - Litsky ML FAU - Farooqui, T AU - Farooqui T FAU - Horrocks, L A AU - Horrocks LA LA - eng ID - NS-10165/NS/NINDS ID - NS-29441/NS/NINDS PT - Journal Article PT - Review PT - Review, Academic CY - UNITED STATES TA - Brain Res Bull JID - 7605818 RN - 0 (Enzyme Inhibitors) RN - 0 (Isoenzymes) RN - EC 3.1.1.- (Phospholipases A) SB - IM MH - Animal MH - Brain Chemistry/*drug effects MH - Carbohydrate Sequence MH - Enzyme Inhibitors/*pharmacology/therapeutic use MH - Human MH - Isoenzymes/antagonists & inhibitors/metabolism MH - Molecular Sequence Data MH - Nervous System Diseases/*drug therapy/enzymology MH - Phospholipases A/*antagonists & inhibitors MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. RF - 142 EDAT- 1999/08/06 MHDA- 1999/08/06 00:01 AID - S0361923099000271 [pii] PST - ppublish SO - Brain Res Bull 1999 Jun;49(3):139-53. UI - 99339460 PMID- 10413115 DA - 19990809 DCOM- 19990809 LR - 20021213 IS - 0014-5793 VI - 454 IP - 1-2 DP - 1999 Jul 2 TI - Phosphorylation of tau protein by recombinant GSK-3beta: pronounced phosphorylation at select Ser/Thr-Pro motifs but no phosphorylation at Ser262 in the repeat domain. PG - 157-64 AB - Glycogen synthase kinase-3beta (GSK-3beta) has been described as a proline-directed kinase which phosphorylates tau protein at several sites that are elevated in Alzheimer paired helical filaments. However, it has been claimed that GSK-3beta can also phosphorylate the non-proline-directed KXGS motifs in the presence of heparin, including Ser262 in the repeat domain of tau, which could induce the detachment of tau from microtubules. We have analyzed the activity of recombinant GSK-3beta and of GSK-3beta preparations purified from tissue, using two-dimensional phosphopeptide mapping, immunoblotting with phosphorylation-sensitive antibodies, and phosphopeptide sequencing. The most prominent phosphorylation sites on tau are Ser396 and Ser404 (PHF-1 epitope), Ser46 and Thr50 in the first insert, followed by a less efficient phosphorylation of other Alzheimer phosphoepitopes (antibodies AT-8, AT-270, etc). We also show that the non-proline-directed activity at KXGS motifs is not due to GSK-3beta itself, but to kinase contaminations in common GSK-3beta preparations from tissues which are activated upon addition of heparin. AD - Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany. mand@mpasmb.desy.de FAU - Godemann, R AU - Godemann R FAU - Biernat, J AU - Biernat J FAU - Mandelkow, E AU - Mandelkow E FAU - Mandelkow, E M AU - Mandelkow EM LA - eng PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Recombinant Proteins) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) SB - IM MH - Animal MH - Ca(2+)-Calmodulin Dependent Protein Kinase/*metabolism MH - Cytoskeleton/enzymology MH - Dose-Response Relationship, Drug MH - Escherichia coli/enzymology MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Heparin/pharmacology MH - Human MH - Immunoblotting MH - Phosphorylation MH - Rabbits MH - Recombinant Proteins MH - Time Factors MH - tau Proteins/*metabolism EDAT- 1999/07/21 MHDA- 1999/07/21 00:01 PST - ppublish SO - FEBS Lett 1999 Jul 2;454(1-2):157-64. UI - 99306826 PMID- 10375392 DA - 19990715 DCOM- 19990715 LR - 20001218 IS - 0003-9861 VI - 367 IP - 1 DP - 1999 Jul 1 TI - Oxidative modification of apolipoprotein E in human very-low-density lipoprotein and its inhibition by glycosaminoglycans. PG - 1-8 AB - The mechanism of metal ion-catalyzed oxidative modification of apolipoprotein E (apoE) in human very-low-density lipoprotein (VLDL) and its inhibition by glycosaminoglycan (GAG) was investigated in vitro. The VLDL oxidation catalyzed by Cu2+ led to the lipid peroxidation, the formation of aggregates, and covalent modification of apoE. The modified apoE lost heparin-binding activity. These results suggest that the lipid peroxidation of VLDL and modification of apoE cause impairment of lipid uptake by cells and deposit the oxidized lipids in the tissues. The lipid peroxidation and oxidative modification of apoE in VLDL mediated by Cu2+ and an aqueous radical generator were suppressed by GAG, heparan sulfate, heparin, and chondroitin sulfate A, even though GAGs demonstrated no ability to scavenge alpha,alpha-diphenyl-beta-picrylhydrazyl radical. There were no relationships between inhibitory activity of GAGs in the VLDL oxidation and their number of sulfate groups which possess chelating activity of metal ion. Therefore, it can be considered that the inhibition of VLDL oxidation by GAGs is possibly due to the interaction between GAG and VLDL which bring about the steric hindrance, interference with the reaction between VLDL particle and the reactive oxygen species. These studies suggest that GAGs preserve the biological functions of apoE from oxidative stress. CI - Copyright 1999 Academic Press. AD - Department of Nutrition, Yamaguchi Prefectural University, 3-2-1 Sakurabatake, Yamaguchi, 753-8502, Japan. FAU - Arai, H AU - Arai H FAU - Kashiwagi, S AU - Kashiwagi S FAU - Nagasaka, Y AU - Nagasaka Y FAU - Uchida, K AU - Uchida K FAU - Hoshii, Y AU - Hoshii Y FAU - Nakamura, K AU - Nakamura K LA - eng PT - Journal Article CY - UNITED STATES TA - Arch Biochem Biophys JID - 0372430 RN - 0 (Aldehydes) RN - 0 (Amidines) RN - 0 (Apolipoproteins E) RN - 0 (Chelating Agents) RN - 0 (Cholesterol Esters) RN - 0 (Free Radical Scavengers) RN - 0 (Glycosaminoglycans) RN - 0 (Lipoproteins, VLDL) RN - 0 (Reactive Oxygen Species) RN - 0 (Thiobarbituric Acid Reactive Substances) RN - 13217-66-8 (2,2'-azobis(2-amidinopropane)) RN - 1898-66-4 (2,2-diphenyl-1-picrylhydrazyl) RN - 2058-59-5 (cholesteryl ester hydroperoxide) RN - 29343-52-0 (4-hydroxy-2-nonenal) RN - 64706-54-3 (Bepridil) RN - 70-18-8 (Glutathione) RN - 7758-98-7 (Copper Sulfate) RN - 9004-54-0 (Dextrans) RN - 9005-49-6 (Heparin) RN - 9007-28-7 (Chondroitin Sulfates) SB - IM MH - Adult MH - Aldehydes/analysis/metabolism MH - Alzheimer Disease MH - Amidines/pharmacology MH - Apolipoproteins E/*metabolism MH - Bepridil/analogs & derivatives/metabolism MH - Chelating Agents/pharmacology MH - Cholesterol Esters/metabolism MH - Chondroitin Sulfates/pharmacology MH - Copper Sulfate/*antagonists & inhibitors/pharmacology MH - Dextrans/pharmacology MH - Free Radical Scavengers/pharmacology MH - Glutathione/pharmacology MH - Glycosaminoglycans/*pharmacology MH - Heparin/analogs & derivatives/metabolism/pharmacology MH - Hippocampus/chemistry MH - Human MH - Hydrogen-Ion Concentration MH - Lipid Peroxidation/*drug effects MH - Lipoproteins, VLDL/*metabolism MH - Male MH - Reactive Oxygen Species/metabolism MH - Thiobarbituric Acid Reactive Substances/metabolism EDAT- 1999/06/22 MHDA- 1999/06/22 00:01 AID - 10.1006/abbi.1999.1222 [doi] AID - S0003986199912225 [pii] PST - ppublish SO - Arch Biochem Biophys 1999 Jul 1;367(1):1-8. UI - 99268614 PMID- 10338292 DA - 19990716 DCOM- 19990716 LR - 20001218 IS - 0306-4522 VI - 90 IP - 4 DP - 1999 TI - Apolipoprotein E promotes the binding and uptake of beta-amyloid into Chinese hamster ovary cells in an isoform-specific manner. PG - 1217-26 AB - The epsilon4 allele of apolipoprotein E gene is a major risk factor for Alzheimer's disease. However, the mechanism by which the E4 isoform of apolipoprotein E increases the risk of Alzheimer's disease is poorly understood. To determine whether the isoform-specific effects of apolipoprotein E may be mediated via clearance of bound beta-amyloid, we examined the uptake of beta-amyloid 1-40 into Chinese hamster ovary cells in the presence or absence of the apolipoprotein E isoforms E2, E3 and E4. Apolipoprotein E2 and E3 treatments were associated with higher association of beta-amyloid with cells as compared to treatment with E4. Heparin blocked the association of beta-amyloid with cells, as did an antibody to one of the apolipoprotein E receptors (the low-density lipoprotein receptor-related protein). Thus, the apolipoproteins E2 and E3, but not E4, may play important roles in the clearance of beta-amyloid from the extracellular space via the low-density lipoprotein receptor-related protein. AD - Sir James McCusker Alzheimer's Disease Research Unit, Hollywood Private Hospital, University of Western Australia, Perth, Australia. FAU - Yang, D S AU - Yang DS FAU - Small, D H AU - Small DH FAU - Seydel, U AU - Seydel U FAU - Smith, J D AU - Smith JD FAU - Hallmayer, J AU - Hallmayer J FAU - Gandy, S E AU - Gandy SE FAU - Martins, R N AU - Martins RN LA - eng PT - Journal Article CY - UNITED STATES TA - Neuroscience JID - 7605074 RN - 0 (Amyloid beta-Protein) RN - 0 (Antibodies) RN - 0 (Apolipoproteins E) RN - 0 (Peptide Fragments) RN - 0 (Receptors, LDL) RN - 0 (amyloid beta-protein (1-40)) RN - 0 (apolipoprotein E-2) RN - 0 (apolipoprotein E-3) RN - 0 (apolipoprotein E-4) RN - 9005-49-6 (Heparin) SB - IM MH - Amyloid beta-Protein/antagonists & inhibitors/*metabolism/pharmacokinetics MH - Animal MH - Antibodies/pharmacology MH - Apolipoproteins E/*pharmacology MH - CHO Cells/*metabolism MH - Hamsters MH - Heparin/pharmacology MH - Peptide Fragments/antagonists & inhibitors/*metabolism/pharmacokinetics MH - Receptors, LDL/immunology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, Non-P.H.S. EDAT- 1999/05/25 MHDA- 1999/05/25 00:01 AID - S0306452298005612 [pii] PST - ppublish SO - Neuroscience 1999;90(4):1217-26. UI - 99264907 PMID- 10332503 DA - 19990623 DCOM- 19990623 LR - 20001218 IS - 1054-3589 VI - 46 DP - 1999 TI - Heparin in inflammation: potential therapeutic applications beyond anticoagulation. PG - 151-208 AB - In this chapter we have described anti-inflammatory functions of heparin distinct from its traditional anticoagulant activity. We have presented in vivo data showing heparin's beneficial effects in various preclinical models of inflammatory disease as well as discussed some clinical studies showing that the anti-inflammatory activities of heparin may translate into therapeutic uses. In vivo models that use low-anticoagulant heparins indicate that the anticoagulant activity can be distinguished from heparin's anti-inflammatory properties. In certain cases such as hypovolemic shock, the efficacy of a low-anticoagulant heparin derivative (GM1892) exceeds heparin. Data also suggest that nonconventional delivery of heparin, specifically via inhalation, has therapeutic potential in improving drug pharmacokinetics (as determined by measuring blood coagulation parameters) and in reducing the persistent concerns of systemic hemorrhagic complications. Results from larger clinical trials with heparin and LMW heparins are eagerly anticipated and will allow us to assess our predictions on the effectiveness of this drug class to treat a variety of human inflammatory diseases. AD - Glycomed Incorporated, Alameda, California 94501, USA. FAU - Tyrrell, D J AU - Tyrrell DJ FAU - Horne, A P AU - Horne AP FAU - Holme, K R AU - Holme KR FAU - Preuss, J M AU - Preuss JM FAU - Page, C P AU - Page CP LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Adv Pharmacol JID - 9015397 RN - 0 (Integrins) RN - 0 (Selectins) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/drug therapy MH - Asthma/drug therapy MH - Colitis, Ulcerative/drug therapy MH - Encephalomyelitis, Experimental Autoimmune/drug therapy MH - Heparin/biosynthesis/pharmacology/*therapeutic use MH - Human MH - Inflammation/*drug therapy/etiology MH - Integrins/physiology MH - Leukocytes/drug effects MH - Platelet Aggregation/drug effects MH - Reperfusion Injury/drug therapy MH - Respiratory Distress Syndrome, Adult/drug therapy MH - Selectins/metabolism MH - Shock/drug therapy RF - 349 EDAT- 1999/05/20 MHDA- 1999/05/20 00:01 PST - ppublish SO - Adv Pharmacol 1999;46:151-208. UI - 99234294 PMID- 10215996 DA - 19990707 DCOM- 19990707 LR - 20001218 IS - 0305-1846 VI - 25 IP - 2 DP - 1999 Apr TI - The amyloid precursor protein of Alzheimer's disease and the Abeta peptide. PG - 81-97 AB - Alzheimer's disease is characterized by the accumulation of beta amyloid peptides in plaques and vessel walls and by the intraneuronal accumulation of paired helical filaments composed of hyperphosphorylated tau. In this review, we concentrate on the biology of amyloid precursor protein, and on the central role of amyloid in the pathogenesis of Alzheimer's disease. Amyloid precursor protein (APP) is part of a super-family of transmembrane and secreted proteins. It appears to have a number of roles, including regulation of haemostasis and mediation of neuroprotection. APP also has potentially important metal and heparin-binding properties, and the current challenge is to synthesize all these varied activities into a coherent view of its function. Cleavage of amyloid precursor protein by beta-and gamma-secretases results in the generation of the Abeta (betaA4) peptide, whereas alpha-secretase cleaves within the Abeta sequence and prevents formation from APP. Recent findings indicate that the site of gamma-secretase cleavage is critical to the development of amyloid deposits; Abeta1-42 is much more amyloidogenic than Abeta1-40. Abeta1-42 formation is favoured by mutations in the two presenilin genes (PS1 and PS2), and by the commonest amyloid precursor protein mutations. Transgenic mouse models of Alzheimer's disease incorporating various mutations in the presenilin gene now exist, and have shown amyloid accumulation and cognitive impairment. Neurofibrillary tangles have not been reproduced in these models, however. While aggregated Abeta is neurotoxic, perhaps via an oxidative mechanism, the relationship between such toxicity and neurofibrillary tangle formation remains a subject of ongoing research. AD - Van Cleef/Roet Centre for Nervous Diseases, Monash University (Alfred Hospital Campus), Prahran, Victoria, Australia. FAU - Storey, E AU - Storey E FAU - Cappai, R AU - Cappai R LA - eng PT - Journal Article PT - Review PT - Review, Academic CY - ENGLAND TA - Neuropathol Appl Neurobiol JID - 7609829 RN - 0 (Amyloid beta-Protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Nerve Tissue Proteins) RN - 0 (Neuroprotective Agents) RN - 0 (Peptide Fragments) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/*metabolism MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Human MH - Molecular Sequence Data MH - Nerve Tissue Proteins/*metabolism MH - Neuroprotective Agents/metabolism MH - Peptide Fragments/*metabolism MH - Structure-Activity Relationship RF - 202 EDAT- 1999/04/24 MHDA- 1999/04/24 00:01 PST - ppublish SO - Neuropathol Appl Neurobiol 1999 Apr;25(2):81-97. UI - 99229757 PMID- 10214944 DA - 19990505 DCOM- 19990505 LR - 20001218 IS - 0014-5793 VI - 447 IP - 2-3 DP - 1999 Mar 26 TI - Accelerated filament formation from tau protein with specific FTDP-17 missense mutations. PG - 195-9 AB - Tau is the major component of the neurofibrillar tangles that are a pathological hallmark of Alzheimers' disease. The identification of missense and splicing mutations in tau associated with the inherited frontotemporal dementia and Parkinsonism linked to chromosome 17 demonstrated that tau dysfunction can cause neurodegeneration. However, the mechanism by which tau dysfunction leads to neurodegeneration remains uncertain. Here, we present evidence that frontotemporal dementia and Parkinsonism linked to chromosome 17 missense mutations, P301L, V337M and R406W, cause an accelerated aggregation of tau into filaments. These results suggest one mechanism by which these mutations can cause neurodegeneration and frontotemporal dementia and Parkinsonism linked to chromosome 17. AD - Department of Pharmacology, Mayo Clinic Jacksonville, FL 32224, USA. FAU - Nacharaju, P AU - Nacharaju P FAU - Lewis, J AU - Lewis J FAU - Easson, C AU - Easson C FAU - Yen, S AU - Yen S FAU - Hackett, J AU - Hackett J FAU - Hutton, M AU - Hutton M FAU - Yen, S H AU - Yen SH LA - eng ID - AG01136/AG/NIA ID - NS37143/NS/NINDS PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Biopolymers) RN - 0 (DNA Primers) RN - 0 (Recombinant Proteins) RN - 0 (tau Proteins) RN - 506-32-1 (Arachidonic Acid) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/genetics/metabolism MH - Arachidonic Acid/pharmacology MH - Base Sequence MH - Biopolymers/chemistry/genetics/metabolism MH - Chromosomes, Human, Pair 17/genetics MH - DNA Primers/genetics MH - Dementia/genetics MH - Heparin/pharmacology MH - Human MH - In Vitro MH - Linkage (Genetics) MH - Microscopy, Electron MH - *Mutation, Missense MH - Nerve Degeneration/genetics MH - Neurofibrillary Tangles/metabolism MH - Parkinson Disease/genetics MH - Recombinant Proteins/genetics/metabolism/ultrastructure MH - Support, U.S. Gov't, P.H.S. MH - tau Proteins/*genetics/*metabolism/ultrastructure EDAT- 1999/04/24 MHDA- 1999/04/24 00:01 PST - ppublish SO - FEBS Lett 1999 Mar 26;447(2-3):195-9. UI - 99215582 PMID- 10201399 DA - 19990423 DCOM- 19990423 LR - 20001218 IS - 1072-8368 VI - 6 IP - 4 DP - 1999 Apr TI - Crystal structure of the N-terminal, growth factor-like domain of Alzheimer amyloid precursor protein. PG - 327-31 AB - Amyloid precursor protein (APP) plays a central role in Alzheimer disease. A proteolytic-breakdown product of APP, called beta-amyloid, is a major component of the diffuse and fibrillar deposits found in Alzheimer diseased brains. The normal physiological role of APP remains largely unknown despite much work. A knowledge of its function will not only provide insights into the genesis of the disease but may also prove vital in the development of an effective therapy. Here we describe the 1.8 A resolution crystal structure of the N-terminal, heparin-binding domain of APP (residues 28-123), which is responsible, among other things, for stimulation of neurite outgrowth. The structure reveals a highly charged basic surface that may interact with glycosaminoglycans in the brain and an abutting hydrophobic surface that is proposed to play an important functional role such as dimerization or ligand binding. Structural similarities with cysteine-rich growth factors, taken together with its known growth-promoting properties, suggests the APP N-terminal domain could function as a growth factor in vivo. AD - The Ian Potter Foundation Protein Crystallography Laboratory, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia. FAU - Rossjohn, J AU - Rossjohn J FAU - Cappai, R AU - Cappai R FAU - Feil, S C AU - Feil SC FAU - Henry, A AU - Henry A FAU - McKinstry, W J AU - McKinstry WJ FAU - Galatis, D AU - Galatis D FAU - Hesse, L AU - Hesse L FAU - Multhaup, G AU - Multhaup G FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL FAU - Parker, M W AU - Parker MW LA - eng SI - PDB/1MWP PT - Journal Article CY - UNITED STATES TA - Nat Struct Biol JID - 9421566 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Growth Substances) RN - 0 (Peptide Fragments) RN - 67256-21-7 (Hepatocyte Growth Factor) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/*chemistry/*metabolism MH - Binding Sites MH - Crystallography, X-Ray MH - Growth Substances/chemistry MH - Heparin/metabolism MH - Hepatocyte Growth Factor/chemistry MH - Models, Molecular MH - Molecular Sequence Data MH - Peptide Fragments/chemistry MH - Protein Conformation MH - Sequence Homology, Amino Acid MH - Support, Non-U.S. Gov't EDAT- 1999/04/14 02:03 MHDA- 2001/03/23 10:01 AID - 10.1038/7562 [doi] PST - ppublish SO - Nat Struct Biol 1999 Apr;6(4):327-31. UI - 99196513 PMID- 10098877 DA - 19990413 DCOM- 19990413 LR - 20001218 IS - 0022-3042 VI - 72 IP - 4 DP - 1999 Apr TI - The sulfate moieties of glycosaminoglycans are critical for the enhancement of beta-amyloid protein fibril formation. PG - 1681-7 AB - Our previous studies have demonstrated that perlecan and perlecan-derived glycosaminoglycans (GAGs) not only bind beta-amyloid protein (Abeta) 1-40 and 1-42, but are also potent enhancers of Abeta fibril formation and stabilize amyloid fibrils once formed. However, it was not determined which moieties in perlecan heparan sulfate GAG chains may be responsible for the observed effects and whether other GAGs were also capable of a similar enhancement of Abeta fibril formation as observed with perlecan GAGs. In the present study, thioflavin T fluorometry (over a 1-week period) was used to extend our previous studies and to test the hypothesis that the sulfate moiety is critical for the enhancing effects of heparin/heparan sulfate GAGs on Abeta 1-40 fibrillogenesis. This hypothesis was confirmed when removal of all sulfates from heparin (i.e., completely desulfated N-acetylated heparin) led to a complete loss in the enhancement of Abeta fibrillogenesis as demonstrated in both thioflavin T fluorometry and Congo red staining studies. On the other hand, removal of O-sulfate from heparin (i.e., completely desulfated N-sulfated heparin), and to a lesser extent N-sulfate (i.e., N-desulfated N-acetylated heparin), resulted in only a partial loss of the enhancement of Abeta 1-40 fibril formation. These studies indicate that the sulfate moieties of GAGs are critical for enhancement of Abeta amyloid fibril formation. In addition, other sulfated molecules such as chondroitin-4-sulfate, dermatan sulfate, dextran sulfate, and pentosan polysulfate all significantly enhanced (greater than twofold by 3 days) Abeta amyloid fibril formation. These latter findings indicate that deposition and accumulation of other GAGs at sites of Abeta amyloid deposition in Alzheimer's disease brain may also participate in the enhancement of Abeta amyloidosis. AD - Department of Pathology, University of Washington, Seattle 98195-6480, USA. FAU - Castillo, G M AU - Castillo GM FAU - Lukito, W AU - Lukito W FAU - Wight, T N AU - Wight TN FAU - Snow, A D AU - Snow AD LA - eng ID - AG05136/AG/NIA ID - AG12953/AG/NIA PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein) RN - 0 (Dyes) RN - 0 (Fibrinolytic Agents) RN - 0 (Glycosaminoglycans) RN - 0 (Peptide Fragments) RN - 0 (Sulfates) RN - 0 (amyloid beta-protein (1-40)) RN - 573-58-0 (Congo Red) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/metabolism/pathology MH - Amyloid beta-Protein/*metabolism MH - Amyloidosis/metabolism/pathology MH - Animal MH - Congo Red MH - Dyes MH - Fibrinolytic Agents/metabolism/pharmacology MH - Glycosaminoglycans/*metabolism MH - Heparin/metabolism/pharmacology MH - Neurofibrillary Tangles/*metabolism/pathology MH - Neurons/drug effects/*metabolism/pathology MH - Peptide Fragments/*metabolism MH - Sulfates/metabolism/pharmacology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Swine EDAT- 1999/03/31 MHDA- 1999/03/31 00:01 PST - ppublish SO - J Neurochem 1999 Apr;72(4):1681-7. UI - 99174934 PMID- 10077316 DA - 19990507 DCOM- 19990507 LR - 20001218 IS - 0306-4522 VI - 89 IP - 2 DP - 1999 Mar TI - Heparin injection into the adult rat hippocampus induces seizures in the absence of macroscopic abnormalities. PG - 329-33 AB - The pathological hallmarks of Alzheimer's disease include neurofibrillary tangles, neuropil threads and neuritic plaques. Neurofibrillary tangles and neuropil threads are comprised of paired helical filaments which are themselves composed of a hyperphosphorylated form of the microtubule-associated protein tau. Neuritic plaques are extracellular deposits of aggregated beta amyloid associated with neurites containing hyperphosphorylated tau. The mechanisms by which the neurofibrillary tangles and neuritic plaques develop in Alzhemier's disease are not clear but it is hypothesized that sulphated glycosaminoglycans are important in their formation. This impression is based on the finding that the glycosaminoglycan, heparan sulphate, is found associated with neurofibrillary tangles, neuritic plaques and neuropil threads while dermatan sulphate, chondroitin sulphate and keratan sulphate immunoreactivity is found around neuritic plaques in brains of Alzheimer's disease patients. Furthermore, in vitro studies demonstrate that sulphated glycosaminoglycans such as heparan sulphate and the closely related molecule heparin interact with tau and potentiate its phosphorylation by a number of serine/threonine kinases, reduce its ability to bind to microtubules and induce paired helical filament formation, all properties associated with tau isolated from Alzheimer's disease brain. Thus, we were interested to learn whether intracerebral injection of the sulphated glycosaminoglycan heparin would give rise to alterations in the cytoskeletal protein tau in the rat brain. Although no cytoskeletal changes were observed, to our considerable surprise we found that the intrahippocampal injection of heparin gave rise to seizures. We have investigated this unexpected effect further in vivo and by using in vitro electrophysiological techniques. AD - University Department of Pharmacology, Oxford, UK. FAU - Mudher, A K AU - Mudher AK FAU - Mellanby, J AU - Mellanby J FAU - McMath, H AU - McMath H FAU - Perry, V H AU - Perry VH FAU - Greene, J R AU - Greene JR LA - eng PT - Journal Article CY - UNITED STATES TA - Neuroscience JID - 7605074 RN - 0 (Drug Combinations) RN - 0 (Quinoxalines) RN - 0 (Receptors, Neurotransmitter) RN - 118876-58-7 (2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline) RN - 485-49-4 (Bicuculline) RN - 9005-49-6 (Heparin) SB - IM MH - Animal MH - Bicuculline/pharmacology MH - Drug Combinations MH - Electrophysiology MH - Heparin/*administration & dosage MH - Hippocampus/pathology/*physiology MH - In Vitro MH - Male MH - Neurons/physiology MH - Quinoxalines/pharmacology MH - Rats MH - Rats, Wistar MH - Receptors, Neurotransmitter/antagonists & inhibitors MH - Seizures/*chemically induced MH - Support, Non-U.S. Gov't MH - Swine MH - Synapses/drug effects/physiology EDAT- 1999/03/17 MHDA- 1999/03/17 00:01 AID - S0306452298005338 [pii] PST - ppublish SO - Neuroscience 1999 Mar;89(2):329-33. UI - 99175182 PMID- 10075702 DA - 19990415 DCOM- 19990415 LR - 20001218 IS - 0021-9258 VI - 274 IP - 12 DP - 1999 Mar 19 TI - Heparin-induced conformational change in microtubule-associated protein Tau as detected by chemical cross-linking and phosphopeptide mapping. PG - 8029-38 AB - In Alzheimer's disease, microtubule-associated protein tau becomes abnormally phosphorylated and aggregates into paired helical filaments. Sulfated glycosaminoglycans such as heparin and heparan sulfate were shown to accumulate in pretangle neurons, stimulate in vitro tau phosphorylation, and cause tau aggregation into paired helical filament-like filaments. The sulfated glycosaminoglycan-tau interaction was suggested to be the central event in the development of neuropathology in Alzheimer's disease brain (Goedert, M., Jakes, R., Spillantini, M. G., Hasegawa, M., Smith, M. J., and Crowther, R. A. (1996) Nature 383, 550-553). The biochemical mechanism by which sulfated glycosaminoglycans stimulate tau phosphorylation and cause tau aggregation remains unclear. In this study, disuccinimidyl suberate (DSS), a bifunctional chemical cross-linker, cross-linked tau dimers, tetramers, high molecular size aggregates, and two tau species of sizes 72 and 83 kDa in the presence of heparin. In the absence of heparin only dimeric tau was cross-linked by DSS. Fast protein liquid chromatography gel filtration revealed that 72- and 83-kDa species were formed by intramolecular cross-linking of tau by DSS. These observations indicate that heparin, in addition to causing aggregation, also induces a conformational change in tau in which reactive groups are unmasked or move closer leading to the DSS cross-linking of 72- and 83-kDa species. Heparin-induced structural changes in tau molecule depended on time of heparin exposure. Dimerization and tetramerization peaked at 48 h, whereas conformational change was completed within 30 min of heparin exposure. Heparin exposure beyond 48 h caused an abrupt aggregation of tau into high molecular size species. Heparin stimulated tau phosphorylation by neuronal cdc2-like kinase (NCLK) and cAMP-dependent protein kinase. Phosphopeptide mapping and phosphopeptide sequencing revealed that tau is phosphorylated by NCLK on Thr212 and Thr231 and by cAMP-dependent protein kinase on Ser262 only in the presence of heparin. Heparin stimulation of tau phosphorylation by NCLK showed dependence on time of heparin exposure and correlated with the heparin-induced conformational change of tau. Our data suggest that heparin-induced conformational change exposes new sites for phosphorylation within tau molecule. AD - Bloomfield Center for Research in Aging, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, and Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec H3T 1E2, Canada. FAU - Paudel, H K AU - Paudel HK FAU - Li, W AU - Li W LA - eng PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Cross-Linking Reagents) RN - 0 (Phosphopeptides) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Amino Acid Sequence MH - Cell Line MH - Chromatography, High Pressure Liquid MH - Cross-Linking Reagents/pharmacology MH - Escherichia coli MH - Heparin/*pharmacology MH - Human MH - Molecular Sequence Data MH - Peptide Mapping MH - Phosphopeptides/chemistry MH - Protein Conformation/drug effects MH - Support, Non-U.S. Gov't MH - tau Proteins/*chemistry/drug effects EDAT- 1999/03/13 MHDA- 1999/03/13 00:01 PST - ppublish SO - J Biol Chem 1999 Mar 19;274(12):8029-38. UI - 99165750 PMID- 10064733 DA - 19990413 DCOM- 19990413 LR - 20001218 IS - 0022-2275 VI - 40 IP - 3 DP - 1999 Mar TI - Competition of Abeta amyloid peptide and apolipoprotein E for receptor-mediated endocytosis. PG - 447-55 AB - The genetic polymorphism of apolipoprotein E (apoE) is associated with the age of onset and relative risk of Alzheimer's disease (AD). In contrast to apoE3, the wild type allele, apoE4 confers an increased risk of late-onset AD. We demonstrate that the beta-amyloid peptide isoforms Abeta (1-28), Abeta (1-40), and Abeta (1-43) compete for the cellular metabolism of apoE3 and apoE4 containing beta-very low density lipoproteins. An antibody raised against Abeta (1-28) cross-reacted with recombinant apoE. Epitope mapping revealed positive amino acid clusters as common epitopes of Abeta (13 through 17; HHQKL) and apoE (residues 144 through 148; LRKRL), both regions known to be heparin binding domains. Abeta in which amino acids 13 through 17 (HHQKL) were replaced by glycine (GGQGL) failed to compete with the cellular uptake of apoE enriched betaVLDL.These observations indicate that Abeta and apoE are taken up into cells by a common pathway involving heparan sulfate proteoglycans. AD - Department of Clinical Chemistry, Albert Ludwigs-University, Freiburg, Germany. FAU - Winkler, K AU - Winkler K FAU - Scharnagl, H AU - Scharnagl H FAU - Tisljar, U AU - Tisljar U FAU - Hoschutzky, H AU - Hoschutzky H FAU - Friedrich, I AU - Friedrich I FAU - Hoffmann, M M AU - Hoffmann MM FAU - Huttinger, M AU - Huttinger M FAU - Wieland, H AU - Wieland H FAU - Marz, W AU - Marz W LA - eng PT - Journal Article CY - UNITED STATES TA - J Lipid Res JID - 0376606 RN - 0 (Amyloid beta-Protein) RN - 0 (Antibodies) RN - 0 (Apolipoproteins E) RN - 0 (Lipoproteins, VLDL) RN - 0 (Membrane Glycoproteins) RN - 0 (Peptide Fragments) RN - 0 (Receptors, Cell Surface) RN - 0 (amyloid beta protein (1-28)) RN - 0 (amyloid beta-protein (1-43)) RN - 0 (apolipoprotein E-3) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/immunology/*metabolism MH - Antibodies/metabolism MH - Apolipoproteins E/*genetics/metabolism MH - Binding, Competitive MH - Cross Reactions/immunology MH - Endocytosis/*physiology MH - Epitope Mapping MH - Fibroblasts MH - Heparitin Sulfate/metabolism MH - Human MH - Lipoproteins, VLDL/metabolism MH - Membrane Glycoproteins/metabolism MH - Molecular Sequence Data MH - Peptide Fragments/immunology/*metabolism/pharmacology MH - Polymorphism (Genetics)/genetics MH - Receptors, Cell Surface/*metabolism MH - Risk Factors MH - Support, Non-U.S. Gov't EDAT- 1999/03/05 MHDA- 1999/03/05 00:01 PST - ppublish SO - J Lipid Res 1999 Mar;40(3):447-55. UI - 99155073 PMID- 10037469 DA - 19990318 DCOM- 19990318 LR - 20001218 IS - 0022-3042 VI - 72 IP - 3 DP - 1999 Mar TI - Mechanism of thrombin clearance by human astrocytoma cells. PG - 980-7 AB - Astroglial cells secrete a variety of factors that contribute to the regulation of neurite initiation and continued outgrowth, among them proteases and protease inhibitors. An alteration in the balance between these proteins has been implicated in Alzheimer's disease, resulting in an accumulation of thrombin:protease nexin 1 (PN1) complexes in the brains of these patients. This report aims at providing a biochemical explanation for this phenomenon. We show that human astrocytoma cells bind and internalize thrombin and thrombin:PN1 complexes efficiently by a PN1-dependent mechanism. Binding was potently inhibited by soluble heparin and did not occur with the mutant PN1 (K7E) deficient in heparin binding. Receptor-associated protein, an antagonist of the low-density lipoprotein receptor-related protein (LRP), inhibited internalization of thrombin by the astrocytoma cells, but did not affect cell-surface binding. The results are consistent with a mechanism by which astrocytoma cells clear thrombin in a sequential manner: thrombin is first complexed with PN1, then bound to cell-surface heparins, and finally internalized by LRP. This mechanism provides a link between the neuronal growth regulators thrombin and PN1 and proteins genetically associated with Alzheimer's disease, such as LRP. AD - Department of Neurology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA. FAU - Mentz, S AU - Mentz S FAU - de Lacalle, S AU - de Lacalle S FAU - Baerga-Ortiz, A AU - Baerga-Ortiz A FAU - Knauer, M F AU - Knauer MF FAU - Knauer, D J AU - Knauer DJ FAU - Komives, E A AU - Komives EA LA - eng ID - AG1204/AG/NIA ID - R01 GM34001/GM/NIGMS ID - R01 HL47463/HL/NHLBI PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Carrier Proteins) RN - 0 (Neoplasm Proteins) RN - 0 (Receptors, Cell Surface) RN - 0 (Receptors, LDL) RN - 0 (Thrombomodulin) RN - 0 (heparin receptor) RN - 0 (protease-nexin) RN - 62229-50-9 (Epidermal Growth Factor) RN - 9005-49-6 (Heparin) RN - EC 3.4.21.5 (Thrombin) SB - IM MH - Astrocytoma/*metabolism/pathology MH - Brain Neoplasms/*metabolism/pathology MH - Carrier Proteins/metabolism MH - Epidermal Growth Factor/metabolism MH - Heparin/metabolism/pharmacology MH - Human MH - Neoplasm Proteins/metabolism MH - Receptors, Cell Surface/metabolism MH - Receptors, LDL/metabolism MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Thrombin/*metabolism MH - Thrombomodulin/metabolism MH - Tumor Cells, Cultured EDAT- 1999/02/26 MHDA- 1999/02/26 00:01 PST - ppublish SO - J Neurochem 1999 Mar;72(3):980-7. UI - 99150407 PMID- 10026239 DA - 19990311 DCOM- 19990311 LR - 20001218 IS - 0022-1554 VI - 47 IP - 3 DP - 1999 Mar TI - Binding and selective detection of the secretory N-terminal domain of the alzheimer amyloid precursor protein on cell surfaces. PG - 373-82 AB - The secretory N-terminal domain of the Alzheimer amyloid precursor protein (sAPP) evokes specific responses in cells on binding to their surfaces. Because APP is expressed in a large variety of cell types, the localization of sAPP binding requires detection techniques that selectively recognize sAPP as a ligand. For this purpose, we prepared antibodies against recombinant sAPP695 (sAPPrec) previously expressed in E. coli. Such antibodies were found to distinguish between sAPPrec and cellular APP or sAPP, as shown by immunocytochemistry and by immunoblot. In addition, they allowed the selective localization of bound sAPPrec on cell surfaces without any signal from cellular APP or sAPP. Saturation of sAPPrec binding to cell surfaces, as determined radiometrically, was reached at 10 nM [125I]-sAPPrec. Binding was specific because it was almost completely inhibited by a 100-fold excess of unlabeled sAPPrec. This specificity of binding was confirmed by surface plasmon resonance spectroscopy. Binding of sAPPrec to cell surfaces occurred in patches and was dependent on the state of cell differentiation. The sAPPrec used in this study contains heparin binding sites, but enzymatic removal of cell surface associated heparin did not affect sAPPrec binding. Aldehyde fixation of cells strongly inhibited their ability to bind sAPPrec. The data point to a fixation-sensitive sAPPrec binding protein which is detectable in the form of patches and therefore is part of assembled cell surface microdomains. AD - Institut fur Zellbiologie and Bonner Forum Biomedizin, Universitat Bonn, Bonn, Germany. FAU - Hoffmann, J AU - Hoffmann J FAU - Pietrzik, C U AU - Pietrzik CU FAU - Kummer, M P AU - Kummer MP FAU - Twiesselmann, C AU - Twiesselmann C FAU - Bauer, C AU - Bauer C FAU - Herzog, V AU - Herzog V LA - eng PT - Journal Article CY - UNITED STATES TA - J Histochem Cytochem JID - 9815334 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Antibodies) RN - 0 (Recombinant Proteins) RN - 50-00-0 (Formaldehyde) RN - 71-00-1 (Histidine) RN - 9005-49-6 (Heparin) RN - EC 4.2.2.7 (Heparin Lyase) SB - IM MH - Amyloid beta-Protein Precursor/immunology/*metabolism MH - Animal MH - Antibodies MH - Biotinylation MH - Cell Line MH - Formaldehyde/pharmacology MH - Heparin/physiology MH - Heparin Lyase/pharmacology MH - Histidine/metabolism MH - Immunohistochemistry MH - Microscopy, Confocal MH - PC12 Cells MH - Protein Binding/drug effects MH - Rats MH - Recombinant Proteins/immunology/metabolism MH - Support, Non-U.S. Gov't EDAT- 1999/02/20 MHDA- 1999/02/20 00:01 PST - ppublish SO - J Histochem Cytochem 1999 Mar;47(3):373-82. UI - 99137830 PMID- 9952400 DA - 19990225 DCOM- 19990225 LR - 20001218 IS - 0270-6474 VI - 19 IP - 4 DP - 1999 Feb 15 TI - Reg1ulatory role and molecular interactions of a cell-surface heparan sulfate proteoglycan (N-syndecan) in hippocampal long-term potentiation. PG - 1226-35 AB - The cellular mechanisms responsible for synaptic plasticity involve interactions between neurons and the extracellular matrix. Heparan sulfates (HSs) constitute a group of glycosaminoglycans that accumulate in the beta-amyloid deposits in Alzheimer's disease and influence the development of neuron-target contacts by interacting with other cell surface and matrix molecules. However, the contribution of HSs to brain function is unknown. We found that HSs play a crucial role in long-term potentiation (LTP), a finding that is consistent with the idea that converging molecular mechanisms are used in the development of neuron-target contacts and in activity-induced synaptic plasticity in adults. Enzymatic cleavage of HS by heparitinase as well as addition of soluble heparin-type carbohydrates prevented expression of LTP in response to 100 Hz/1 sec stimulation of Schaffer collaterals in rat hippocampal slices. A prominent carrier protein for the type of glycans implicated in LTP regulation in the adult hippocampus was identified as N-syndecan (syndecan-3), a transmembrane proteoglycan that was expressed at the processes of the CA1 pyramidal neurons in an activity-dependent manner. Addition of soluble N-syndecan into the CA1 dendritic area prevented tetanus-induced LTP. A major substrate of src-type kinases, cortactin (p80/85), and the tyrosine kinase fyn copurified with N-syndecan from hippocampus. Moreover, association of both cortactin and fyn to N-syndecan was rapidly increased after induction of LTP. N-syndecan may thus act as an important regulator in the activity-dependent modulation of neuronal connectivity by transmitting signals between extracellular heparin-binding factors and the fyn signaling pathway. AD - Department of Biosciences, Division of Animal Physiology, 00014 University of Helsinki, Helsinki, Finland. FAU - Lauri, S E AU - Lauri SE FAU - Kaukinen, S AU - Kaukinen S FAU - Kinnunen, T AU - Kinnunen T FAU - Ylinen, A AU - Ylinen A FAU - Imai, S AU - Imai S FAU - Kaila, K AU - Kaila K FAU - Taira, T AU - Taira T FAU - Rauvala, H AU - Rauvala H LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurosci JID - 8102140 RN - 0 (Membrane Glycoproteins) RN - 0 (Microfilament Proteins) RN - 0 (Proteoglycans) RN - 0 (cortactin) RN - 0 (syndecan 3) RN - EC 4.2.2. (Polysaccharide-Lyases) RN - EC 4.2.2.8 (heparitinsulfate lyase) SB - IM MH - Animal MH - Cells, Cultured MH - Electric Stimulation MH - Electrophysiology MH - Hippocampus/*drug effects/ultrastructure MH - Immunoblotting MH - In Situ Hybridization MH - In Vitro MH - Long-Term Potentiation/*drug effects MH - Male MH - Membrane Glycoproteins/*pharmacology MH - Membrane Potentials/physiology MH - Microfilament Proteins/metabolism MH - Patch-Clamp Techniques MH - Polysaccharide-Lyases/pharmacology MH - Proteoglycans/*pharmacology MH - Pyramidal Cells/drug effects MH - Rats MH - Rats, Wistar MH - Support, Non-U.S. Gov't MH - Synapses/*drug effects EDAT- 1999/02/10 MHDA- 1999/02/10 00:01 PST - ppublish SO - J Neurosci 1999 Feb 15;19(4):1226-35. UI - 98332547 PMID- 9665729 DA - 19980812 DCOM- 19980812 LR - 20001218 IS - 0006-2960 VI - 37 IP - 28 DP - 1998 Jul 14 TI - Rapid assembly of Alzheimer-like paired helical filaments from microtubule-associated protein tau monitored by fluorescence in solution. PG - 10223-30 AB - Alzheimer's disease is characterized by the progressive deposition of two types of fibers in the affected brains, the amyloid fibers (consisting of the Abeta peptide, generating the amyloid plaques) and paired helical filaments (PHFs, made up of tau protein, forming the neurofibrillary tangles). While the principles of amyloid aggregation are known in some detail, the investigation of PHF assembly has been hampered by the low efficiency of tau aggregation, the requirement of high protein concentrations, and the lack of suitable detection methods. Here we report a quantitative assay system that permits monitoring of the assembly of PHFs in real time by the fluorescence of dyes such as thioflavine S or T. Using this assay, we evaluated parameters that influence the efficiency of filament formation. Disulfide-linked dimers of tau constructs representing the repeat domain assemble into PHFs most efficiently, but other tau isoforms or constructs form bona fide PHFs as well. The rate of assembly is greatly enhanced by polyanions such as RNA, heparin, and notably polyglutamate which resembles the acidic tail of tubulin. The assembly is optimal at pH approximately 6 and low ionic strengths (<50 mM) and increases steeply with temperatures above 30 degreesC, indicating that it is an entropy-driven process. AD - Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany. FAU - Friedhoff, P AU - Friedhoff P FAU - Schneider, A AU - Schneider A FAU - Mandelkow, E M AU - Mandelkow EM FAU - Mandelkow, E AU - Mandelkow E LA - eng PT - Journal Article CY - UNITED STATES TA - Biochemistry JID - 0370623 RN - 0 (Buffers) RN - 0 (Fluorescent Dyes) RN - 0 (Polymers) RN - 0 (Salts) RN - 0 (Solutions) RN - 0 (Thiazoles) RN - 0 (polyanions) RN - 0 (tau Proteins) RN - 2390-54-7 (thioflavin T) SB - IM MH - Alzheimer Disease/*metabolism MH - Buffers MH - Fluorescent Dyes/chemistry/metabolism MH - Human MH - Hydrogen-Ion Concentration MH - Neurofibrillary Tangles/*chemistry/*metabolism/ultrastructure MH - Polymers/chemistry/metabolism MH - *Protein Structure, Secondary MH - Salts MH - Solutions MH - Spectrometry, Fluorescence MH - Support, Non-U.S. Gov't MH - Temperature MH - Thiazoles/chemistry/metabolism MH - tau Proteins/*chemistry/*metabolism/ultrastructure EDAT- 1998/07/17 MHDA- 1998/07/17 00:01 AID - 10.1021/bi980537d [doi] AID - bi980537d [pii] PST - ppublish SO - Biochemistry 1998 Jul 14;37(28):10223-30. UI - 98210104 PMID- 9541497 DA - 19980512 DCOM- 19980512 LR - 20001218 IS - 0021-9738 VI - 101 IP - 8 DP - 1998 Apr 15 TI - Phenotype-dependent differences in apolipoprotein E metabolism and in cholesterol homeostasis in human monocyte-derived macrophages. PG - 1670-7 AB - In this study, we investigated the impact of the common apoE polymorphism on apoE metabolism and cholesterol homeostasis in monocyte-derived macrophages isolated from E2/2, E3/3, and E4/4 subjects. Unloaded cells of all genotypes contained similar amounts of free cholesterol, cholesteryl ester, and apoE mRNA. E3/3 cells secreted 77 and 30% more apoE than E2/2 or E4/4 cells, respectively. Pulse-chase studies confirmed that the apoE secretion rate was greatest in E3/3 and least in E2/2 cells and showed that a portion of apoE2, but not apoE3 or apoE4, was degraded intracellularly. Surface binding of apoE was greatest in E4/4 cells, as revealed by heparinase treatment. On cholesterol loading with acetylated LDL, apoE mRNA levels and protein secretion rose most in E4/4 and least in E2/2 cells. Cholesterol and cholesteryl ester content, however, rose most in E2/2 and least in E3/3 cells. Incubations with 3H-cholesterol-labeled acetylated LDL revealed that E2/2 cells were most efficient at secreting cholesterol. The greatest reuptake of 3H-cholesterol-rich particles was from E4/4 macrophage- conditioned media. Thus, E2/2 macrophages, despite a low apoE secretion rate, are protected from cholesterol storage by apoE-mediated cholesterol efflux. In E3/3 macrophages, cholesterol accumulation is lessened by a high basal apoE secretion rate. E4/4 macrophages secrete the most apoE but lack effective net cholesterol efflux due to enhanced surface binding and reuptake of cholesterol-rich particles. AD - Institut fur Arterioskleroseforschung, Westfalische Wilhelms-Universitat, 48149 Munster, Germany. cullen@uni-muenster.de FAU - Cullen, P AU - Cullen P FAU - Cignarella, A AU - Cignarella A FAU - Brennhausen, B AU - Brennhausen B FAU - Mohr, S AU - Mohr S FAU - Assmann, G AU - Assmann G FAU - von Eckardstein, A AU - von Eckardstein A LA - eng PT - Journal Article CY - UNITED STATES TA - J Clin Invest JID - 7802877 RN - 0 (Apolipoproteins E) RN - 0 (Cholesterol Esters) RN - 0 (RNA, Messenger) RN - 145-63-1 (Suramin) RN - 57-88-5 (Cholesterol) RN - EC 4.2.2.7 (Heparin Lyase) SB - AIM SB - IM MH - Adult MH - Alzheimer Disease/etiology/genetics/metabolism MH - Apolipoproteins E/*genetics/*metabolism/physiology MH - Arteriosclerosis/etiology/genetics/metabolism MH - Cholesterol/*metabolism MH - Cholesterol Esters/metabolism MH - Female MH - Foam Cells/metabolism MH - Gene Expression MH - Heparin Lyase/pharmacology MH - Homeostasis MH - Human MH - In Vitro MH - Macrophages/drug effects/*metabolism/physiology MH - Male MH - Middle Age MH - Monocytes/metabolism MH - Phenotype MH - RNA, Messenger/genetics/metabolism MH - Support, Non-U.S. Gov't MH - Suramin/pharmacology EDAT- 1998/05/16 MHDA- 1998/05/16 00:01 PST - ppublish SO - J Clin Invest 1998 Apr 15;101(8):1670-7. UI - 98239233 PMID- 9579630 DA - 19980706 DCOM- 19980706 LR - 20010323 IS - 0094-6176 VI - 24 IP - 2 DP - 1998 TI - Thrombin and antithrombotics. PG - 87-91 AB - From injury through healing, thrombin has several important functions in blood clotting, subsequent clot lysis, and tissue repair. These include edema, inflammation, cell recruitment, cellular releases, transformations, mitogenesis, and angiogenesis. Thrombin also participates in disease states, such as venous thrombosis, coronary thrombosis, stroke, and pulmonary emboli, among others and is implicated in atherosclerosis, the growth and metastasis of certain cancers, Alzheimer's disease, and perhaps other conditions. Thrombin must be continually generated to sustain normal and pathogenic processes. This is because of a variety of consumptive mechanisms. Unlike other activated factors in thrombotic and fibrinolytic pathways, and because thrombin promotes its own generation (feedback and cellular activation), thrombin is a primary target for therapeutics. Besides recombinant hirudins, Argatroban (Novastan) and Bivalirudin (Hirulog) are promising thrombin-directed inhibitors for antithrombotic intervention. AD - New York State Department of Health, Albany 12201, USA. FAU - Fenton, J W 2nd AU - Fenton JW 2nd FAU - Ofosu, F A AU - Ofosu FA FAU - Brezniak, D V AU - Brezniak DV FAU - Hassouna, H I AU - Hassouna HI LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Semin Thromb Hemost JID - 0431155 RN - 0 (Blood Coagulation Factors) RN - 0 (Coumarins) RN - 0 (Fibrinolytic Agents) RN - 12001-79-5 (Vitamin K) RN - 8001-27-2 (Hirudin) RN - 9001-26-7 (Prothrombin) RN - 9001-27-8 (Factor VIII) RN - 9005-49-6 (Heparin) RN - 9035-58-9 (Thromboplastin) RN - 91-64-5 (coumarin) RN - EC 3.4.21.5 (Thrombin) SB - IM MH - Blood Coagulation/drug effects/physiology MH - Blood Coagulation Factors/physiology MH - Coumarins/pharmacology MH - Enzyme Activation MH - Factor VIII/physiology MH - Fibrinolytic Agents/*pharmacology MH - Heparin/pharmacology/therapeutic use MH - Hirudin/pharmacology MH - Human MH - Prothrombin/metabolism MH - Thrombin/*physiology MH - Thromboplastin/physiology MH - Vitamin K/physiology RF - 48 EDAT- 1998/05/14 02:04 MHDA- 2001/03/28 10:01 PST - ppublish SO - Semin Thromb Hemost 1998;24(2):87-91. UI - 98206749 PMID- 9546672 DA - 19980513 DCOM- 19980513 LR - 20021213 IS - 0014-2956 VI - 252 IP - 3 DP - 1998 Mar 15 TI - Sequential phosphorylation of Tau by glycogen synthase kinase-3beta and protein kinase A at Thr212 and Ser214 generates the Alzheimer-specific epitope of antibody AT100 and requires a paired-helical-filament-like conformation. PG - 542-52 AB - AT100 is a monoclonal antibody highly specific for phosphorylated Tau in Alzheimer paired helical filaments. Here we show that the epitope is generated by a complex sequence of sequential phosphorylation, first of Ser199, Ser202 and Thr205 (around the epitope of antibody AT8), next of Thr212 by glycogen synthase kinase (GSK)-3beta (a proline-directed kinase), then of Ser214 by protein kinase A (PKA). Conversely, if Ser214 is phosphorylated first it protects Thr212 and the Ser-Pro motifs around the AT8 site against phosphorylation, and the AT100 epitope is not formed. The generation of the AT100 epitope requires a conformation of tau induced by polyanions such as heparin, RNA or poly(Glu), conditions which also favor the formation of paired helical filaments. The Alzheimer-like phosphorylation can be induced by brain extracts. In the extract, the kinases responsible for generating the AT100 epitope are GSK-3beta and PKA, which can be inhibited by their specific inhibitors LiCl and RII, respectively. A cellular model displaying the reaction with AT100 is presented by Sf9 insect cells transfected with Tau. Knowledge of the events and kinases generating the AT100 epitope in cells might allow us to study the degeneration of the cytoskeleton in Alzheimer's disease. AD - Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany. FAU - Zheng-Fischhofer, Q AU - Zheng-Fischhofer Q FAU - Biernat, J AU - Biernat J FAU - Mandelkow, E M AU - Mandelkow EM FAU - Illenberger, S AU - Illenberger S FAU - Godemann, R AU - Godemann R FAU - Mandelkow, E AU - Mandelkow E LA - eng PT - Journal Article CY - GERMANY TA - Eur J Biochem JID - 0107600 RN - 0 (Amyloid beta-Protein) RN - 0 (Antibodies, Monoclonal) RN - 0 (Epitopes) RN - 0 (Phosphopeptides) RN - 0 (Recombinant Proteins) RN - 0 (tau Proteins) RN - 56-45-1 (Serine) RN - 72-19-5 (Threonine) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Cyclic AMP-Dependent Protein Kinases) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/chemistry/immunology MH - Antibodies, Monoclonal MH - Brain/*metabolism MH - Ca(2+)-Calmodulin Dependent Protein Kinase/*metabolism MH - Cloning, Molecular MH - Cyclic AMP-Dependent Protein Kinases/metabolism MH - *Epitopes MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Human MH - Molecular Sequence Data MH - Mutagenesis, Site-Directed MH - Peptide Mapping MH - Phosphopeptides/chemistry MH - Phosphorylation MH - Point Mutation MH - *Protein Structure, Secondary MH - Recombinant Proteins/chemistry/immunology/metabolism MH - *Serine MH - Substrate Specificity MH - *Threonine MH - tau Proteins/*chemistry/immunology/*metabolism EDAT- 1998/04/18 MHDA- 1998/04/18 00:01 PST - ppublish SO - Eur J Biochem 1998 Mar 15;252(3):542-52. UI - 98140958 PMID- 9473690 DA - 19980402 DCOM- 19980402 LR - 20001218 IS - 0006-8993 VI - 779 IP - 1-2 DP - 1998 Jan 1 TI - Hepatocyte growth factor (HGF/SF) in Alzheimer's disease. PG - 262-70 AB - Hepatocyte growth factor (HGF/SF), is a heparin-binding polypeptide which stimulates DNA synthesis in a variety of cell types and also promotes cell migration and morphogenesis. HGF/SF mRNA has been found in a variety of tissues, including brain. In a previous study, we showed that basic fibroblast growth factor (bFGF), another heparin-binding protein is increased in Alzheimer's disease (AD), and appears to be associated with the heparan-sulfate proteoglycans bound to B/A4 amyloid (Biochem. Biophys. Res. Commun. 171 (1990) 690-696). In the present study, we examined the distribution of HGF/SF in 4% paraformaldehyde fixed samples of prefrontal cortex from control and Alzheimer patients, in order to assess the possibility that HGF/SF may be found in association with the pathologic changes which occur in Alzheimer's disease. A specific polyclonal antibody directed against HGF/SF revealed widespread HGF/SF-like immunoreactivity in both the cerebral cortex and white matter. Confocal microscopy confirmed that HGF/SF could be found in both GFAP positive astrocytes and LN3 positive microglia cells, as well as rare scattered cortical neurons. In the AD cases studied, the immunoreactivity was increased within both the astrocytes and microglial cells surrounding individual senile plaques. No staining was seen within the neurofibrillary tangles. Western blot analysis confirmed the normal molecular form of HGF/SF in Alzheimer's disease. Quantitative ELISA assay demonstrated a significant increase in HGF/SF in AD relative to age matched controls. These studies confirm the presence of HGF/SF immunoreactivity within neurons, astrocytes and microglial cells. They also indicate that HGF/SF may be increased within senile plaques as a function of the gliosis and microglial proliferation which occurs in association with these structures in Alzheimer's disease. AD - Department of Pathology, Brown University School of Medicine/Rhode Island Hospital, Providence 02903, USA. FAU - Fenton, H AU - Fenton H FAU - Finch, P W AU - Finch PW FAU - Rubin, J S AU - Rubin JS FAU - Rosenberg, J M AU - Rosenberg JM FAU - Taylor, W G AU - Taylor WG FAU - Kuo-Leblanc, V AU - Kuo-Leblanc V FAU - Rodriguez-Wolf, M AU - Rodriguez-Wolf M FAU - Baird, A AU - Baird A FAU - Schipper, H M AU - Schipper HM FAU - Stopa, E G AU - Stopa EG LA - eng ID - AG10682/AG/NIA ID - MH/NS 31862/MH/NIMH PT - Journal Article CY - NETHERLANDS TA - Brain Res JID - 0045503 RN - 0 (Glial Fibrillary Acidic Protein) RN - 67256-21-7 (Hepatocyte Growth Factor) SB - IM MH - Aged MH - Aged, 80 and over MH - Alzheimer Disease/*metabolism/pathology MH - Astrocytes/chemistry MH - Blotting, Western MH - Case-Control Studies MH - Enzyme-Linked Immunosorbent Assay MH - Fluorescent Antibody Technique MH - Glial Fibrillary Acidic Protein/analysis MH - Hepatocyte Growth Factor/*analysis MH - Human MH - Immunohistochemistry MH - Microscopy, Confocal MH - Middle Age MH - Prefrontal Cortex/*chemistry/pathology MH - Support, U.S. Gov't, P.H.S. EDAT- 1998/02/25 MHDA- 1998/02/25 00:01 PST - ppublish SO - Brain Res 1998 Jan 1;779(1-2):262-70. UI - 98058954 PMID- 9395501 DA - 19980115 DCOM- 19980115 LR - 20001218 IS - 0021-9258 VI - 272 IP - 50 DP - 1997 Dec 12 TI - Heparin-binding properties of the amyloidogenic peptides Abeta and amylin. Dependence on aggregation state and inhibition by Congo red. PG - 31617-24 AB - Aggregation and deposition of the 40-42-residue amyloid beta-protein (Abeta) are early and necessary neuropathological events in Alzheimer's disease. An understanding of the molecular interactions that trigger these events is important for therapeutic strategies aimed at blocking Abeta plaque formation at the earliest stages. Heparan sulfate proteoglycans may play a fundamental role since they are invariably associated with Abeta and other amyloid deposits at all stages. However, the nature of the Abeta-heparan sulfate proteoglycan binding has been difficult to elucidate because of the strong tendency of Abeta to self-aggregate. Affinity co-electrophoresis can measure the binding of proteoglycans or glycosaminoglycans to proteins without altering the physical state of the protein during the assay. We used affinity co-electrophoresis to study the interaction between Abeta and the glycosaminoglycan heparin and found that the aggregation state of Abeta governs its heparin-binding properties: heparin binds to fibrillar but not nonfibrillar Abeta. The amyloid binding dye, Congo red, inhibited the interaction in a specific and dose-dependent manner. The "Dutch" mutant AbetaE22Q peptide formed fibrils more readily than wild type Abeta and it also attained a heparin-binding state more readily, but, once formed, mutant and wild type fibrils bound heparin with similar affinities. The heparin-binding ability of aggregated AbetaE22Q was reversible with incubation in a solvent that promotes alpha-helical conformation, further suggesting that conformation of the peptide is important. Studies with another human amyloidogenic protein, amylin, suggested that its heparin-binding properties were also dependent on aggregation state. These results demonstrate the dependence of the Abeta-heparin interaction on the conformation and aggregation state of Abeta rather than primary sequence alone, and suggest methods of interfering with this association. AD - Department of Neurology and Program in Neuroscience, Harvard Medical School, and Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA. FAU - Watson, D J AU - Watson DJ FAU - Lander, A D AU - Lander AD FAU - Selkoe, D J AU - Selkoe DJ LA - eng ID - AG 06173/AG/NIA ID - AG 12749/AG/NIA ID - NS 26862/NS/NINDS PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein) RN - 0 (Dyes) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta-protein (1-40)) RN - 106602-62-4 (amylin) RN - 573-58-0 (Congo Red) RN - 9005-49-6 (Heparin) SB - IM MH - Amino Acid Sequence MH - Amyloid/*metabolism MH - Amyloid beta-Protein/genetics/*metabolism MH - Animal MH - Cerebral Amyloid Angiopathy/genetics/metabolism MH - Congo Red/*pharmacology MH - Dyes/*pharmacology MH - Electrophoresis/methods MH - Heparin/*metabolism MH - Human MH - Kinetics MH - Molecular Sequence Data MH - Peptide Fragments/metabolism MH - Protein Binding MH - Rats MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1998/02/12 MHDA- 1998/02/12 00:01 PST - ppublish SO - J Biol Chem 1997 Dec 12;272(50):31617-24. UI - 98114314 PMID- 9453569 DA - 19980224 DCOM- 19980224 LR - 20001218 IS - 0022-3042 VI - 70 IP - 2 DP - 1998 Feb TI - Heparin oligosaccharides that pass the blood-brain barrier inhibit beta-amyloid precursor protein secretion and heparin binding to beta-amyloid peptide. PG - 736-44 AB - We have previously demonstrated that full-length heparin stimulates the synthesis and secretion of beta-amyloid precursor protein (APP) through an amyloidogenic pathway in neuroblastoma cells. In the present study, heparin was chemically depolymerized, and the effect of low-molecular-weight (LMW) heparin on APP secretion was investigated. In contrast to full-length heparin, LMW heparin had no significant effect on APP secretion. However, LMW heparin fragments, especially heparin disaccharides, were able to inhibit efficiently the stimulatory effect of heparin on APP secretion. LMW heparin derivatives also inhibit the binding of heparin to the beta-amyloid peptide (1-28). Using an in vitro model, we further demonstrated the passage of LMW heparin derivatives through the blood-brain barrier. This study suggests that LMW heparin derivatives or analogues may be effective as therapeutic agents to prevent or slow the process of amyloidogenesis in Alzheimer's disease. AD - Henry L. Schwartz Department of Geriatrics and Adult Development, Mount Sinai Medical Center, New York, New York, USA. FAU - Leveugle, B AU - Leveugle B FAU - Ding, W AU - Ding W FAU - Laurence, F AU - Laurence F FAU - Dehouck, M P AU - Dehouck MP FAU - Scanameo, A AU - Scanameo A FAU - Cecchelli, R AU - Cecchelli R FAU - Fillit, H AU - Fillit H LA - eng ID - NS35092/NS/NINDS PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Heparin, Low-Molecular-Weight) RN - 0 (Oligosaccharides) RN - 9005-49-6 (Heparin) SB - IM MH - Amyloid beta-Protein Precursor/*biosynthesis MH - Animal MH - Astrocytes/cytology/*physiology MH - Blood-Brain Barrier/drug effects/*physiology MH - Capillaries MH - Cerebrovascular Circulation MH - Coculture MH - Endothelium, Vascular/cytology/*physiology MH - Heparin/chemistry/*metabolism MH - Heparin, Low-Molecular-Weight/*pharmacology MH - Kinetics MH - Models, Neurological MH - Oligosaccharides/pharmacokinetics/*pharmacology MH - Rats MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1998/02/07 MHDA- 1998/02/07 00:01 PST - ppublish SO - J Neurochem 1998 Feb;70(2):736-44. UI - 98070514 PMID- 9407097 DA - 19980123 DCOM- 19980123 LR - 20021213 IS - 0021-9258 VI - 272 IP - 52 DP - 1997 Dec 26 TI - Alzheimer-like changes in microtubule-associated protein Tau induced by sulfated glycosaminoglycans. Inhibition of microtubule binding, stimulation of phosphorylation, and filament assembly depend on the degree of sulfation. PG - 33118-24 AB - Hyperphosphorylated microtubule-associated protein tau is the major proteinaceous component of the paired helical and straight filaments which constitute a defining neuropathological characteristic of Alzheimer's disease and a number of other neurodegenerative disorders. We have recently shown that full-length recombinant tau assembles into Alzheimer-like filaments upon incubation with heparin. Heparin also promotes phosphorylation of tau by a number of protein kinases, prevents tau from binding to taxol-stabilized microtubules, and produces rapid disassembly of microtubules assembled from tau and tubulin. Here, we have used the above parameters to study the interactions between tau protein and a number of naturally occurring and synthetic glycosaminoglycans. We show that the magnitude of the glycosaminoglycan effects is proportional to their degree of sulfation. Thus, the strongly sulfated glycosaminoglycans dextran sulfate, pentosan polysulfate, and heparin were the most potent, whereas the non-sulfated dextran and hyaluronic acid were without effect. The moderately sulfated glycosaminoglycans heparan sulfate, chondroitin sulfate, and dermatan sulfate had intermediate effects, whereas keratan sulfate had little or no effect. These in vitro interactions between tau protein and sulfated glycosaminoglycans reproduced the known characteristics of paired helical filament-tau from Alzheimer's disease brain. Sulfated glycosaminoglycans are present in nerve cells in Alzheimer's disease brain in the early stages of neurofibrillary degeneration, suggesting that their interactions with tau may constitute a central event in the development of the neuronal pathology of Alzheimer's disease. AD - Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom. FAU - Hasegawa, M AU - Hasegawa M FAU - Crowther, R A AU - Crowther RA FAU - Jakes, R AU - Jakes R FAU - Goedert, M AU - Goedert M LA - eng PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Glycosaminoglycans) RN - 0 (Sulfates) RN - 0 (tau Proteins) RN - 63231-63-0 (RNA) RN - 9005-49-6 (Heparin) RN - 9007-49-2 (DNA) RN - 9050-30-0 (Heparitin Sulfate) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Protein-Serine-Threonine Kinases) RN - EC 2.7.10.- (cyclin-dependent kinase 5) SB - IM MH - Alzheimer Disease/*metabolism MH - Ca(2+)-Calmodulin Dependent Protein Kinase/metabolism MH - DNA/metabolism MH - Glycogen Synthase Kinase 3 MH - Glycosaminoglycans/*metabolism MH - Heparin/metabolism MH - Heparitin Sulfate/metabolism MH - Human MH - Microtubules/*metabolism MH - Neurofibrillary Tangles/*metabolism MH - Phosphorylation MH - Protein Conformation MH - Protein-Serine-Threonine Kinases/metabolism MH - RNA/metabolism MH - Sulfates/*metabolism MH - Support, Non-U.S. Gov't MH - tau Proteins/*metabolism EDAT- 1998/01/31 MHDA- 1998/01/31 00:01 PST - ppublish SO - J Biol Chem 1997 Dec 26;272(52):33118-24. UI - 98041526 PMID- 9375678 DA - 19971212 DCOM- 19971212 LR - 20001218 IS - 0022-3042 VI - 69 IP - 6 DP - 1997 Dec TI - Perlecan binds to the beta-amyloid proteins (A beta) of Alzheimer's disease, accelerates A beta fibril formation, and maintains A beta fibril stability. PG - 2452-65 AB - Perlecan is a specific heparan sulfate proteoglycan that accumulates in the fibrillar beta-amyloid (A beta) deposits of Alzheimer's disease. Perlecan purified from the Engelbreth-Holm-Swarm tumor was used to define perlecan's interactions with A beta and its effects on A beta fibril formation. Using a solid-phase binding immunoassay, freshly solubilized full-length A beta peptides bound immobilized perlecan at two sites, representing both high-affinity [K(D) = approximately 5.8 x 10(-11) M for A beta (1-40); K(D) = approximately 6.5 x 10(-12) M for A beta (1-42)] and lower-affinity [K(D) = 3.5 x 10(-8) M for A beta (1-40); K(D) = 4.3 x 10(-8) M for A beta (1-42)] interactions. An increase in the binding capacity of A beta (1-40) to perlecan correlated with an increase in A beta amyloid fibril formation during a 1-week incubation period. The high-capacity binding of A beta (1-40) to perlecan was similarly observed using perlecan heparan sulfate glycosaminoglycans and was completely abolished by heparin, but not by chondroitin-4-sulfate. Using a thioflavin T fluorometry assay, perlecan accelerated the rate of A beta (1-40) amyloid fibril formation, causing a significant increase in A beta fibril assembly over a 2-week incubation period at 1 h (2.8-fold increase), 1 day (3.6-fold increase), and 3 days (2.8-fold increase) in comparison with A beta (1-40) alone. Perlecan also initially accelerated the formation of A beta (1-42) fibrils within 1 h and maintained significantly higher levels of A beta (1-42) thioflavin T fluorescence throughout a 2-week experimental period in comparison with A beta (1-42) alone, suggesting perlecan's ability to maintain amyloid fibril stability. Perlecan's effects on A beta (1-40) fibril formation and maintenance of A beta (1-42) fibril stability occurred in a dose-dependent manner and was also mediated primarily by perlecan's glycosaminoglycan chains. Perlecan was the most effective enhancer and accelerator of A beta fibril formation when compared directly with other amyloid plaque components, including apolipoprotein E, alpha1-antichymotrypsin, P component, C1q, and C3. This study, therefore, demonstrates that perlecan not only binds to the predominant isoforms of A beta, but also accelerates A beta fibril formation and stabilizes amyloid fibrils once formed, confirming pivotal roles for perlecan in the pathogenesis of A beta amyloidosis in Alzheimer's disease. AD - Neuropathology Laboratories, University of Washington, Seattle 98195-6480, U.S.A. FAU - Castillo, G M AU - Castillo GM FAU - Ngo, C AU - Ngo C FAU - Cummings, J AU - Cummings J FAU - Wight, T N AU - Wight TN FAU - Snow, A D AU - Snow AD LA - eng ID - AG05136/AG/NIA ID - AG12953/AG/NIA ID - HL18645/HL/NHLBI PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein) RN - 0 (Glycosaminoglycans) RN - 0 (Peptide Fragments) RN - 0 (Proteoglycans) RN - 0 (amyloid beta-protein (1-40)) RN - 0 (beta-amyloid (1-42)) RN - 143972-95-6 (perlecan) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein/*drug effects/*metabolism/physiology MH - Binding, Competitive MH - Glycosaminoglycans/physiology MH - Heparitin Sulfate/chemistry/*metabolism/*pharmacology/physiology MH - Peptide Fragments/metabolism MH - Proteoglycans/chemistry/*metabolism/*pharmacology MH - Support, U.S. Gov't, P.H.S. MH - Time Factors EDAT- 1997/12/31 MHDA- 1997/12/31 00:01 PST - ppublish SO - J Neurochem 1997 Dec;69(6):2452-65. UI - 98019020 PMID- 9357988 DA - 19971125 DCOM- 19971125 LR - 20001218 IS - 0014-5793 VI - 415 IP - 3 DP - 1997 Oct 6 TI - Expression and analysis of heparin-binding regions of the amyloid precursor protein of Alzheimer's disease. PG - 303-7 AB - Deletion mutagenesis studies have suggested that there are two domains within APP which bind heparan sulphate. These domains have been cloned and expressed in the yeast Pichia pastoris. Both recombinant proteins bound to heparin. One domain (APP316-447) was further characterised by binding studies with peptides encompassing this region. Peptides homologous to APP316-346 and APP416-447 were found to bind heparin. Circular dichroism studies show that APP416-447 shifted towards an alpha-helical conformation in the presence of heparin. This study suggests that heparin-binding domains may lie within regions high in alpha-helical structure. AD - Department of Pathology, The University of Melbourne, Parkville, Victoria, Australia. FAU - Mok, S S AU - Mok SS FAU - Sberna, G AU - Sberna G FAU - Heffernan, D AU - Heffernan D FAU - Cappai, R AU - Cappai R FAU - Galatis, D AU - Galatis D FAU - Clarris, H J AU - Clarris HJ FAU - Sawyer, W H AU - Sawyer WH FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL FAU - Small, D H AU - Small DH LA - eng PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Peptide Fragments) RN - 0 (Recombinant Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease MH - Amyloid beta-Protein Precursor/chemistry/*genetics MH - Binding Sites MH - Blotting, Western MH - Chromatography, Affinity MH - Circular Dichroism MH - Cloning, Molecular MH - Escherichia coli/genetics MH - Gene Expression MH - Heparin/*metabolism MH - Human MH - Mutagenesis MH - Peptide Fragments/chemistry/genetics/metabolism MH - Protein Binding MH - *Protein Structure, Secondary MH - Recombinant Proteins/chemistry/metabolism MH - Sequence Deletion MH - Support, Non-U.S. Gov't EDAT- 1997/11/14 MHDA- 1997/11/14 00:01 AID - S0014579397011460 [pii] PST - ppublish SO - FEBS Lett 1997 Oct 6;415(3):303-7. UI - 98002689 PMID- 9341206 DA - 19971117 DCOM- 19971117 LR - 20001218 IS - 0006-2960 VI - 36 IP - 43 DP - 1997 Oct 28 TI - Interaction between glycosaminoglycans and immunoglobulin light chains. PG - 13187-94 AB - Amyloidosis is a pathological process in which normally soluble proteins polymerize to form insoluble fibrils (amyloid). Amyloid formation is found in a number of diseases, including Alzheimer's disease, adult-onset diabetes, and light-chain-associated amyloidosis. No pharmaceutical methods currently exist to prevent this process or to remove the fibrils from tissue. The search for treatment and prevention methods is hampered by a limited understanding of the biophysical basis of amyloid formation. Glycosaminoglycans (GAGs) are long, unbranched heteropolysaccharides composed of repeating disaccharide subunits and are known to associate with amyloid fibrils. The interaction of amyloid-associated free light chains with GAGs was tested by both size-exclusion high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis experiments. The results indicated that heparin 16 000 and chondroitin sulfate B and C precipitated both human intact light chains and recombinant light chain variable domains. Although all light chains interacted with heparin, the strongest interactions were obtained with proteins that had formed amyloid. Molecular modeling indicated the possibility of interaction between heparin and the conserved saddlelike surface of the light chain dimer opposite the complementarity-determining segments that form part of the antigen-binding site of a functional antibody. This suggestion might offer a new path to block the aggregation of amyloid-associated light chain proteins, by design of antagonists based on properties of GAG binding. A hexasaccharide was modeled as the basis for a possible antagonist. AD - Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, Argonne, Illinois 60439, USA. FAU - Jiang, X AU - Jiang X FAU - Myatt, E AU - Myatt E FAU - Lykos, P AU - Lykos P FAU - Stevens, F J AU - Stevens FJ LA - eng ID - DK43757/DK/NIDDK PT - Journal Article CY - UNITED STATES TA - Biochemistry JID - 0370623 RN - 0 (Glycosaminoglycans) RN - 0 (Immunoglobulin Variable Region) RN - 0 (Immunoglobulins, Light-Chain) RN - 0 (Recombinant Proteins) SB - IM MH - Amyloidosis/metabolism MH - Chromatography, High Pressure Liquid MH - Dimerization MH - Electrophoresis, Polyacrylamide Gel MH - Glycosaminoglycans/*metabolism MH - Human MH - Immunoglobulin Variable Region/chemistry/genetics/metabolism MH - Immunoglobulins, Light-Chain/chemistry/genetics/*metabolism MH - Models, Molecular MH - Multiple Myeloma/metabolism MH - Protein Structure, Tertiary MH - Recombinant Proteins/metabolism MH - Support, U.S. Gov't, Non-P.H.S. MH - Support, U.S. Gov't, P.H.S. EDAT- 1997/10/28 MHDA- 1997/10/28 00:01 AID - 10.1021/bi970408h [doi] AID - bi970408h [pii] PST - ppublish SO - Biochemistry 1997 Oct 28;36(43):13187-94. UI - 97368224 PMID- 9221767 DA - 19971003 DCOM- 19971003 LR - 20011102 IS - 0270-6474 VI - 17 IP - 15 DP - 1997 Aug 1 TI - Neurotoxicity of the 22 kDa thrombin-cleavage fragment of apolipoprotein E and related synthetic peptides is receptor-mediated. PG - 5678-86 AB - Potent neurotoxicity is associated with both apolipoprotein E (apoE)-related synthetic peptides and the 22 kDa N-terminal thrombin-cleavage fragment of apoE. Furthermore, the E4 isoform of the 22 kDa fragment is significantly more toxic than the same fragment derived from the E3 isoform, suggesting the possibility of a direct role of apoE-associated neurotoxicity in the pathophysiology of Alzheimer's disease. In the present study, the potential role of cell surface receptors in mediating neurotoxicity was assessed by using a variety of agents that should block the heparin-binding and receptor-binding activity of apoE. Effective inhibitors of neurotoxicity of both the apoE peptides and the apoE fragment include heparin, heparan sulfate, sodium chlorate and heparinase, the low-density lipoprotein (LDL) receptor-related protein receptor-associated protein, and a polyclonal anti-LDL receptor-related protein antibody. These results suggest that the neurotoxicity of the 22 kDa thrombin cleavage fragment of apoE and related peptides is receptor-mediated, and that the most likely candidate receptor is a heparan sulfate proteoglycan-LDL receptor-related protein complex. AD - Department of Neurosurgery, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA. FAU - Tolar, M AU - Tolar M FAU - Marques, M A AU - Marques MA FAU - Harmony, J A AU - Harmony JA FAU - Crutcher, K A AU - Crutcher KA LA - eng ID - HL27333/HL/NHLBI ID - NS31410/NS/NINDS PT - Journal Article CY - UNITED STATES TA - J Neurosci JID - 8102140 RN - 0 (Apolipoproteins E) RN - 0 (LDL-Receptor Related Protein 1) RN - 0 (Receptors, Lipoprotein) RN - 9050-30-0 (Heparitin Sulfate) RN - EC 3.4.21.5 (Thrombin) SB - IM MH - Animal MH - Apolipoproteins E/*toxicity MH - Cell Death/*drug effects MH - Cells, Cultured/drug effects MH - Chick Embryo MH - Dose-Response Relationship, Drug MH - Heparitin Sulfate/*toxicity MH - LDL-Receptor Related Protein 1 MH - Receptors, Lipoprotein/*drug effects MH - Support, U.S. Gov't, P.H.S. MH - Thrombin/pharmacology EDAT- 1997/08/01 MHDA- 1997/08/01 00:01 PST - ppublish SO - J Neurosci 1997 Aug 1;17(15):5678-86. UI - 97297526 PMID- 9152995 DA - 19970707 DCOM- 19970707 LR - 20001218 IS - 0197-0186 VI - 30 IP - 6 DP - 1997 Jun TI - Heparin promotes beta-secretase cleavage of the Alzheimer's amyloid precursor protein. PG - 543-8 AB - In Alzheimer's disease, abnormal processing of the amyloid precursor protein (APP) is thought to play an important role in amyloid deposition. We investigated the effect of heparin, a highly sulfated glycosaminoglycan related to heparan sulfate, on the secretion of the beta-secretase cleavage product of APP (sAPP beta) in a human neuroblastoma cell line. Heparin induced an increase in the secretion of total APP, and an even greater relative increase in the secretion of sAPP beta. The effect on sAPP beta was specific to heparin. These data support the hypothesis that highly sulfated heparan sulfate proteoglycans may promote amyloidogenic pathways of APP metabolism. AD - Faculty of Pharmacy, University of Alberta, Edmonton, Canada. leveugle@gpu.srv.ualberta.ca FAU - Leveugle, B AU - Leveugle B FAU - Ding, W AU - Ding W FAU - Durkin, J T AU - Durkin JT FAU - Mistretta, S AU - Mistretta S FAU - Eisle, J AU - Eisle J FAU - Matic, M AU - Matic M FAU - Siman, R AU - Siman R FAU - Greenberg, B D AU - Greenberg BD FAU - Fillit, H M AU - Fillit HM LA - eng PT - Journal Article CY - ENGLAND TA - Neurochem Int JID - 8006959 RN - 0 (Amyloid beta-Protein Precursor) RN - 9005-49-6 (Heparin) RN - EC 3.4.- (Endopeptidases) RN - EC 3.4.99.- (secretase) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein Precursor/biosynthesis/*metabolism/secretion MH - Endopeptidases/*metabolism MH - Heparin/*pharmacology MH - Human MH - Neuroblastoma/metabolism MH - Support, Non-U.S. Gov't MH - Tumor Cells, Cultured EDAT- 1997/06/01 MHDA- 1997/06/01 00:01 AID - S0197018696001039 [pii] PST - ppublish SO - Neurochem Int 1997 Jun;30(6):543-8. UI - 97281766 PMID- 9136074 DA - 19970619 DCOM- 19970619 LR - 20001218 IS - 0730-2312 VI - 65 IP - 2 DP - 1997 May TI - Interaction between Alzheimer's disease beta A4 precursor protein (APP) and the extracellular matrix: evidence for the participation of heparan sulfate proteoglycans. PG - 145-58 AB - The interaction between the Alzheimer amyloid precursor protein (APP) and an intact extracellular matrix (ECM), matrigel, obtained from Engelbreth-Holm-Swarm tumors was evaluated. Based on quantitative analyses of the binding data obtained from solid phase binding assays, two binding sites on the ECM were identified for [125I]-APP (with apparent Kd1 of 1.0 x 10(-11) M and Kd2 of 1.6 x 10(-9) M respectively). Over 70% of [125I]-APP was displaced by heparin and N-desulfated heparin but not by chondroitin sulfate. Pretreatment of matrigel with heparitinase decreased the binding of [125I]-APP by 80%. beta-amyloid peptides (residues 1-40, 1-28, and 1-16) containing a heparin binding domain also displaced 80% of bound [125I]-APP, which was totally displaced by intact APP. The binding of [125I]-APP to matrigel increased by 210% with a decrease in the pH. These observations suggest that [125I]-APP interacts mainly with heparan sulfate proteoglycan present in the ECM. The binding of [125I]-APP to individual ECM components was also analyzed. [125I]-APP was found to bind laminin and collagen type IV but not fibronectin. However, when these ECM constituents were combined, the extent of APP-binding decreased significantly, to levels comparable to those obtained with intact matrigel, suggesting that multiple interactions may occur between ECM constituents and [125I]-APP. The results are discussed in terms of APP function and amyloidogenesis. AD - Departamento de Biologia Celular y Molecular, Facultad de Ciencias Biologicas, P. Universidad Catolica de Chile, Santiago, Chile. FAU - Caceres, J AU - Caceres J FAU - Brandan, E AU - Brandan E LA - eng PT - Journal Article CY - UNITED STATES TA - J Cell Biochem JID - 8205768 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Drug Combinations) RN - 0 (Fibronectins) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Iodine Radioisotopes) RN - 0 (Laminin) RN - 0 (Proteoglycans) RN - 119978-18-6 (matrigel) RN - 9007-34-5 (Collagen) RN - 9050-30-0 (Heparitin Sulfate) RN - EC 4.2.2. (Polysaccharide-Lyases) RN - EC 4.2.2.8 (heparitinsulfate lyase) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein Precursor/*metabolism MH - Binding Sites MH - Binding, Competitive MH - Collagen/metabolism MH - Drug Combinations MH - Extracellular Matrix/*metabolism MH - Fibronectins/metabolism MH - Heparan Sulfate Proteoglycan MH - Heparitin Sulfate/*metabolism MH - Human MH - Hydrogen-Ion Concentration MH - Iodine Radioisotopes MH - Kinetics MH - Laminin/metabolism MH - Polysaccharide-Lyases/pharmacology MH - Proteoglycans/*metabolism MH - Support, Non-U.S. Gov't EDAT- 1997/05/01 MHDA- 2000/06/20 09:00 AID - 10.1002/(SICI)1097-4644(199705)65:2<145::AID-JCB2>3.0.CO;2-U [pii] PST - ppublish SO - J Cell Biochem 1997 May;65(2):145-58. UI - 97200934 PMID- 9048763 DA - 19970407 DCOM- 19970407 LR - 20001218 IS - 0022-3042 VI - 68 IP - 3 DP - 1997 Mar TI - Identification of heparin-binding domains in the amyloid precursor protein of Alzheimer's disease by deletion mutagenesis and peptide mapping. PG - 1164-72 AB - Recent studies have shown that the binding of the amyloid protein precursor (APP) of Alzheimer's disease to heparan sulfate proteoglycans (HSPGs) can modulate a neurite outgrowth-promoting function associated with APP. We used three different approaches to identify heparin-binding domains in APP. First, as heparin-binding domains are likely to be within highly folded regions of proteins, we analyzed the secondary structure of APP using several predictive algorithms. This analysis showed that two regions of APP695 contain a high degree of secondary structure, and clusters of basic residues, considered mandatory for heparin binding, were found, principally within these regions. To determine which domains of APP bind heparin, deletion mutants of APP695 were prepared and analyzed for binding to a heparin affinity column. The results suggested that there must be at least two distinct heparin-binding regions in APP. To identify novel heparin-binding regions, peptides homologous to candidate heparin-binding domains were analyzed for their ability to bind heparin. These experiments suggested that APP contains at least four heparin-binding domains. The presence of more than one heparin-binding domain on APP suggests the possibility that APP may interact with more than one type of glycosaminoglycan. AD - Department of Pathology, University of Melbourne, Parkville, Victoria, Australia. FAU - Clarris, H J AU - Clarris HJ FAU - Cappai, R AU - Cappai R FAU - Heffernan, D AU - Heffernan D FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL FAU - Small, D H AU - Small DH LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein Precursor) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*genetics/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/chemistry/*genetics/*metabolism MH - Gene Deletion MH - Glycosylation MH - Heparin/*metabolism MH - Human MH - Molecular Sequence Data MH - Mutagenesis MH - Peptide Mapping MH - Protein Structure, Secondary MH - Support, Non-U.S. Gov't EDAT- 1997/03/01 MHDA- 1997/03/01 00:01 PST - ppublish SO - J Neurochem 1997 Mar;68(3):1164-72. UI - 97094206 PMID- 8940294 DA - 19970103 DCOM- 19970103 LR - 20001218 IS - 0892-6638 VI - 10 IP - 13 DP - 1996 Nov TI - Human apolipoprotein E: the Alzheimer's disease connection. PG - 1485-94 AB - Human apolipoprotein (apo) E, long known for its prominent role in cholesterol transport and plasma lipoprotein metabolism, has recently emerged as a major genetic risk factor for Alzheimer's disease, a neurodegenerative disorder. In a variety of populations worldwide, one of the three common alleles of apoE, apoE4, is overrepresented in Alzheimer's subjects compared with age- and sex-matched controls. The genetic and epidemiologic evidence suggests that apoE is a major susceptibility gene for Alzheimer's disease; it likely accounts for a major portion of the genetic heterogeneity in the disease. Although its role in the development of Alzheimer's disease is unknown, biochemical and cell biology studies are providing important insights into how apoE may be involved in neurodegenerative disorders. Based on an understanding of the structure and function of apoE in lipid transport and cellular metabolism, it is suggested that apoE is involved in a final common pathway of neuronal repair and remodeling: apoE3 (most common allele) supporting effective repair and remodeling after neuronal injury by noxious agents, and apoE4 being less effective in these processes. AD - Gladstone Institute of Cardiovascular Disease, University of California at San Francisco, 94141-9100, USA. FAU - Weisgraber, K H AU - Weisgraber KH FAU - Mahley, R W AU - Mahley RW LA - eng ID - HL41633/HL/NHLBI PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - FASEB J JID - 8804484 RN - 0 (Apolipoproteins E) RN - 0 (Proteoglycans) RN - 0 (Receptors, LDL) RN - 0 (heparin proteoglycan) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*etiology/genetics/metabolism MH - Apolipoproteins E/chemistry/genetics/*physiology MH - Cytoskeleton/metabolism MH - Genotype MH - Heparin/analogs & derivatives/metabolism MH - Human MH - Neurons/metabolism MH - Protein Conformation MH - Proteoglycans/metabolism MH - Receptors, LDL/metabolism MH - Structure-Activity Relationship MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - tau Proteins/metabolism RF - 58 EDAT- 1996/11/01 MHDA- 1996/11/01 00:01 PST - ppublish SO - FASEB J 1996 Nov;10(13):1485-94. UI - 97016880 PMID- 8863493 DA - 19961202 DCOM- 19961202 LR - 20001218 IS - 0022-3042 VI - 67 IP - 5 DP - 1996 Nov TI - Increased activity-regulating and neuroprotective efficacy of alpha-secretase-derived secreted amyloid precursor protein conferred by a C-terminal heparin-binding domain. PG - 1882-96 AB - Proteolytic cleavage of beta-amyloid precursor protein (beta APP) by alpha-secretase results in release of one secreted form (sAPP) of APP (sAPP alpha), whereas cleavage by beta-secretase releases a C-terminally truncated sAPP (sAPP beta) plus amyloid beta-peptide (A beta). beta APP mutations linked to some inherited forms of Alzheimer's disease may alter its processing such that levels of sAPP alpha are reduced and levels of sAPP beta increased. sAPP alpha s may play important roles in neuronal plasticity and survival, whereas A beta can be neurotoxic. sAPP alpha was approximately 100-fold more potent than sAPP beta in protecting hippocampal neurons against excitotoxicity, A beta toxicity, and glucose deprivation. Whole-cell patch clamp and calcium imaging analyses showed that sAPP beta was less effective than sAPP alpha in suppressing synaptic activity, activating K+ channels, and attenuating calcium responses to glutamate. Using various truncated sAPP alpha and sAPP beta APP695 products generated by eukaryotic and prokaryotic expression systems, and synthetic sAPP peptides, the activity of sAPP alpha was localized to amino acids 591-612 at the C-terminus. Heparinases greatly reduced the actions of sAPP alpha s, indicating a role for a heparin-binding domain at the C-terminus of sAPP alpha in receptor activation. These findings indicate that alternative processing of beta APP has profound effects on the bioactivity of the resultant sAPP products and suggest that reduced levels of sAPP alpha could contribute to neuronal degeneration in Alzheimer's disease. AD - Sanders-Brown Research Center on Aging, University of Kentucky, Lexington 40536-0230, USA. FAU - Furukawa, K AU - Furukawa K FAU - Sopher, B L AU - Sopher BL FAU - Rydel, R E AU - Rydel RE FAU - Begley, J G AU - Begley JG FAU - Pham, D G AU - Pham DG FAU - Martin, G M AU - Martin GM FAU - Fox, M AU - Fox M FAU - Mattson, M P AU - Mattson MP LA - eng ID - AG10917/AG/NIA ID - NS29001/NS/NINDS ID - NS30583/NS/NINDS PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Peptide Fragments) RN - 0 (Potassium Channels) RN - 0 (Receptors, AMPA) RN - 0 (Receptors, Kainic Acid) RN - 0 (Receptors, N-Methyl-D-Aspartate) RN - 0 (Recombinant Fusion Proteins) RN - 0 (amyloid beta-protein (25-35)) RN - 56-86-0 (Glutamic Acid) RN - 7440-70-2 (Calcium) RN - 9005-49-6 (Heparin) RN - EC 2.5.1.18 (Glutathione Transferase) RN - EC 3.4.- (Endopeptidases) RN - EC 3.4.99.- (secretase) RN - EC 4.2.2. (Polysaccharide-Lyases) RN - EC 4.2.2.7 (Heparin Lyase) SB - IM EIN - J Neurochem 1997 Mar;68(3):1331 MH - Alzheimer Disease/genetics/metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/*pharmacology MH - Amyloid beta-Protein Precursor/biosynthesis/chemistry/*metabolism MH - Animal MH - Base Sequence MH - Binding Sites MH - Calcium/metabolism MH - Cell Line MH - Cell Survival/drug effects MH - Cells, Cultured MH - Cloning, Molecular MH - Endopeptidases/*metabolism MH - Escherichia coli MH - Fetus MH - Glutamic Acid/pharmacology MH - Glutathione Transferase MH - Heparin/*metabolism MH - Heparin Lyase MH - Hippocampus/*physiology MH - Human MH - Kidney MH - Molecular Sequence Data MH - Mutagenesis, Site-Directed MH - Neurons/cytology/drug effects/*physiology MH - Patch-Clamp Techniques MH - Peptide Fragments/*pharmacology MH - Polymerase Chain Reaction MH - Polysaccharide-Lyases/pharmacology MH - Potassium Channels/drug effects/physiology MH - Rats MH - Receptors, AMPA/physiology MH - Receptors, Kainic Acid/physiology MH - Receptors, N-Methyl-D-Aspartate/physiology MH - Recombinant Fusion Proteins/biosynthesis/metabolism MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1996/11/01 MHDA- 1996/11/01 00:01 PST - ppublish SO - J Neurochem 1996 Nov;67(5):1882-96. UI - 97002330 PMID- 8849730 DA - 19961112 DCOM- 19961112 LR - 20011126 IS - 0028-0836 VI - 383 IP - 6600 DP - 1996 Oct 10 TI - Assembly of microtubule-associated protein tau into Alzheimer-like filaments induced by sulphated glycosaminoglycans. PG - 550-3 AB - The paired helical filament (PHF) is the major component of the neurofibrillary deposits that form a defining neuropathological characteristic of Alzheimer's disease. PHFs are composed of microtubule-associated protein tau, in a hyperphosphorylated state. Hyperphosphorylation of tau results in its inability to bind to microtubules and is believed to precede PHF assembly. However, it is unclear whether hyperphosphorylation of tau is either necessary or sufficient for PHF formation. Here we show that non-phosphorylated recombinant tau isoforms with three microtubule-binding repeats form paired helical-like filaments under physiological conditions in vitro, when incubated with sulphated glycosaminoglycans such as heparin or heparan sulphate. Furthermore, heparin prevents tau from binding to microtubules and promotes microtubule disassembly. Finally, we show that heparan sulphate and hyperphosphorylated tau coexist in nerve cells of the Alzheimer's disease brain at the earliest known stages of neurofibrillary pathology. These findings, with previous studies which show that heparin stimulates tau phosphorylation by a number of protein kinases, indicate that sulphated glycosaminoglycans may be a key factor in the formation of the neurofibrillary lesions of Alzheimer's disease. AD - MRC Laboratory of Molecular Biology, Cambridge, UK. FAU - Goedert, M AU - Goedert M FAU - Jakes, R AU - Jakes R FAU - Spillantini, M G AU - Spillantini MG FAU - Hasegawa, M AU - Hasegawa M FAU - Smith, M J AU - Smith MJ FAU - Crowther, R A AU - Crowther RA LA - eng PT - Journal Article CY - ENGLAND TA - Nature JID - 0410462 RN - 0 (Recombinant Proteins) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) RN - 9050-30-0 (Heparitin Sulfate) SB - IM CIN - Nature. 1996 Oct 10;383(6600):476-7. PMID: 8849716 MH - Alzheimer Disease/*metabolism/pathology MH - Escherichia coli MH - Heparin/*metabolism MH - Heparitin Sulfate/*metabolism MH - Hippocampus/metabolism/pathology MH - Human MH - Neurofibrils/metabolism/ultrastructure MH - Phosphorylation MH - Protein Binding MH - Recombinant Proteins/metabolism MH - Support, Non-U.S. Gov't MH - tau Proteins/*metabolism/ultrastructure EDAT- 1996/10/10 MHDA- 1996/10/10 00:01 PST - ppublish SO - Nature 1996 Oct 10;383(6600):550-3. UI - 96355760 PMID- 8752125 DA - 19961002 DCOM- 19961002 LR - 20001218 IS - 0022-3042 VI - 67 IP - 3 DP - 1996 Sep TI - Polymerization of tau into filaments in the presence of heparin: the minimal sequence required for tau-tau interaction. PG - 1183-90 AB - Paired helical filaments isolated from the brains of patients with Alzheimer's disease are composed of a major protein component, the microtubule-associated protein termed tau, together with other nonprotein components, including heparan, a glycosaminoglycan, the more extensively sulfated form of which is heparin. As some of these nonprotein components may modulate the assembly of tau into filamentous structures, we have analyzed the ability of the whole tau protein or some of its fragments to self-assemble in the presence of heparin. Different tau fragments, all of them containing some sequences of the tubulin-binding motif, can assemble in vitro into filaments. We have also found formation of polymers with the 18-residue-long peptide corresponding to the third tubulin-binding motif of tau. This suggests that the ability of tau for self-assembly could be localized in a short sequence of amino acids present in the tubulin-binding repeats of the tau molecule. AD - Centro de Biologia Molecular Severo Ochoa, (CSIC-UAM), Universidad Autonoma de Madrid, Spain. FAU - Perez, M AU - Perez M FAU - Valpuesta, J M AU - Valpuesta JM FAU - Medina, M AU - Medina M FAU - Montejo de Garcini, E AU - Montejo de Garcini E FAU - Avila, J AU - Avila J LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Anticoagulants) RN - 0 (Peptide Fragments) RN - 0 (Polymers) RN - 0 (tau Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Amino Acid Sequence MH - Anticoagulants/chemistry/*pharmacology MH - Base Sequence MH - Heparin/chemistry/*pharmacology MH - Human MH - Microfilaments/*chemistry/drug effects/metabolism MH - Molecular Sequence Data MH - Peptide Fragments/chemistry MH - Polymers MH - Protein Structure, Tertiary MH - Support, Non-U.S. Gov't MH - tau Proteins/*chemistry/drug effects/metabolism EDAT- 1996/09/01 MHDA- 1996/09/01 00:01 PST - ppublish SO - J Neurochem 1996 Sep;67(3):1183-90. UI - 96370839 PMID- 8774735 DA - 19961024 DCOM- 19961024 LR - 20001218 IS - 0014-2956 VI - 239 IP - 3 DP - 1996 Aug 1 TI - Ligand-binding sites in human serum amyloid P component. PG - 850-6 AB - Amyloid P component (AP) is a naturally occurring glycoprotein that is found in serum and basement membranes. AP is also a component of all types of amyloid, including that found in individuals who suffer from Alzheimer's disease and Down's syndrome. Because AP has been found to bind strongly and specifically to certain glycosaminoglycans that are components of amyloid deposits, AP may play an important role in the maintenance of amyloid. In the present work, we isolated and identified two proteolytic fragments of AP that are responsible for its heparin-binding activity. Neither fragment corresponds to published heparin-binding sequences. The structural requirements for activity of the peptides (amino acid residues 27-38 and 192-203 of AP) were examined by means of solid-phase inhibition assays with synthetic peptides. AP-(192-203)-peptide inhibits the Ca(2+)-dependent binding of AP to heparin with an IC50 of 25 microM, while the IC50 of AP-(27-38)-peptide and AP-(33-38)-peptide are 10 microM and 2 microM, respectively. The understanding of the structure and function of active AP peptides will be useful for development of amyloid-targeted diagnostics and therapeutics. AD - Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark. FAU - Heegaard, N H AU - Heegaard NH FAU - Heegaard, P M AU - Heegaard PM FAU - Roepstorff, P AU - Roepstorff P FAU - Robey, F A AU - Robey FA LA - eng PT - Journal Article CY - GERMANY TA - Eur J Biochem JID - 0107600 RN - 0 (Amyloid P Component) RN - 0 (Ligands) RN - 0 (Peptide Fragments) RN - 58-85-5 (Biotin) RN - 9005-49-6 (Heparin) RN - EC 3.4.21.4 (Trypsin) SB - IM MH - Amino Acid Sequence MH - Amyloid P Component/drug effects/*metabolism MH - Binding Sites MH - Binding, Competitive MH - Biotin MH - Dose-Response Relationship, Drug MH - Heparin/*metabolism MH - Human MH - Ligands MH - Molecular Sequence Data MH - Peptide Fragments/metabolism MH - Support, Non-U.S. Gov't MH - Trypsin/pharmacology EDAT- 1996/08/01 MHDA- 1996/08/01 00:01 PST - ppublish SO - Eur J Biochem 1996 Aug 1;239(3):850-6. UI - 96313123 PMID- 8756325 DA - 19960923 DCOM- 19960923 LR - 20001218 IS - 1072-8368 VI - 3 IP - 8 DP - 1996 Aug TI - Inhibitory conformation of the reactive loop of alpha 1-antitrypsin. PG - 676-81 AB - The reactive site loop of the serpin family of serine proteinase inhibitors is flexible and can adopt a number of diverse conformations. A 2.9 A resolution structure of alpha 1-antitrypsin-the principal proteinase inhibitor in human plasma-shows the loop in a stable canonical conformation matching that found in all other families of serine proteinase inhibitors. This unexpected finding in the absence of loop insertion into the body of the molecule favours a two-stage mechanism of inhibition and provides a model for the heparin activation of antithrombin. The beta-pleated strand conformation of the loop also accounts for the polymerization of the serpins in disease and for their association with other beta-sheet structures, most notably the beta-amyloid of Alzheimer's disease. AD - Department of Haematology, University of Cambridge, UK. FAU - Elliott, P R AU - Elliott PR FAU - Lomas, D A AU - Lomas DA FAU - Carrell, R W AU - Carrell RW FAU - Abrahams, J P AU - Abrahams JP LA - eng PT - Journal Article CY - UNITED STATES TA - Nat Struct Biol JID - 9421566 RN - 0 (Polymers) RN - 0 (alpha 1-Antitrypsin) RN - 9005-49-6 (Heparin) SB - IM MH - Comparative Study MH - Crystallography, X-Ray MH - Heparin/pharmacology MH - Human MH - Liver Cirrhosis/etiology MH - Models, Molecular MH - Mutation MH - Polymers MH - Protein Conformation MH - Structure-Activity Relationship MH - Support, Non-U.S. Gov't MH - alpha 1-Antitrypsin/*chemistry/drug effects/genetics/ultrastructure EDAT- 1996/08/01 MHDA- 1996/08/01 00:01 PST - ppublish SO - Nat Struct Biol 1996 Aug;3(8):676-81. UI - 96159064 PMID- 8591996 DA - 19960401 DCOM- 19960401 LR - 20001218 IS - 0021-9541 VI - 166 IP - 2 DP - 1996 Feb TI - Extracellular matrix regulates the amount of the beta-amyloid precursor protein and its amyloidogenic fragments. PG - 360-9 AB - We have studied the influence of the extracellular matrix (ECM) on the amount of beta-amyloid precursor protein (APP) and C-terminal amyloid-bearing fragments in 313 fibroblasts. After incubation with ECM components, the cellular APP content of 3T3 cells changed. Besides, different substrata including collagen, fibronectin, laminin, vitronectin, and heparin, determined changes in the amount of a C-terminal 22 kDa-fragment. The regulation of amyloidogenic fragments by the ECM was transient; in fact, when 3T3 cells were plated on tissue culture dishes coated with collagen or vitronectin, maximal levels of the 22 kDa fragment were observed 12 h after plating; in the presence of fibronectin, the maximum level of the amyloidogenic fragment was obtained 36 h after plating. These results indicate that the ECM modulates in a transient way the generation of APP-derived polypeptides containing the amyloid-beta-peptide (A beta). The ECM does not have a generalized effect on 3T3 fibroblasts, because no significant differences in cell attachment, growth rate, whole-cell polypeptide pattern beta 1 integrin and alpha-tubulin levels were observed on cells grown on various matrix proteins. Laminin, collagen, and heparin also influence the level of an amyloidogenic fragment of 35 kDa in Neuro 2A neuronal cells, without a significant change in the neuronal marker acetylcholinesterase. In this case, however, a long-lasting response to ECM molecules was observed. These observations provide evidence that ECM molecules influence APP biogenesis, including the generation of amyloidogenic fragments containing the A beta peptide. Our studies might prove significant to understand the localized increment of beta-amyloid deposition in selected areas of the brain of Alzheimer's patients. AD - Departamento de Biologia Celular y Molecular, Facultad de Ciencias Biologicas, Pontificia Universidad Catolica de Chile, Santiago, Chile. FAU - Bronfman, F C AU - Bronfman FC FAU - Soto, C AU - Soto C FAU - Tapia, L AU - Tapia L FAU - Tapia, V AU - Tapia V FAU - Inestrosa, N C AU - Inestrosa NC LA - eng PT - Journal Article CY - UNITED STATES TA - J Cell Physiol JID - 0050222 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Laminin) RN - 0 (Peptide Fragments) RN - 9005-49-6 (Heparin) RN - 9007-34-5 (Collagen) SB - IM MH - 3T3 Cells MH - Amyloid/*biosynthesis MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Collagen/pharmacology MH - Extracellular Matrix/*physiology MH - Fibroblasts/metabolism MH - Heparin/pharmacology MH - Laminin/pharmacology MH - Mice MH - Neurons/metabolism MH - Peptide Fragments/*metabolism MH - Support, Non-U.S. Gov't MH - Tumor Cells, Cultured EDAT- 1996/02/01 MHDA- 2000/06/20 09:00 AID - 10.1002/(SICI)1097-4652(199602)166:2<360::AID-JCP14>3.0.CO;2-F [pii] PST - ppublish SO - J Cell Physiol 1996 Feb;166(2):360-9. UI - 96290692 PMID- 8730853 DA - 19961125 DCOM- 19961125 LR - 20021101 IS - 0959-4965 VI - 7 IP - 2 DP - 1996 Jan 31 TI - HB-GAM is a cytokine present in Alzheimer's and Down's syndrome lesions. PG - 667-71 AB - The distribution of heparin binding growth associated molecule (HB-GAM) in the cerebral amyloidoses of Alzheimer's disease (AD) and Down's syndrome (DS), conditions characterized by the deposition of amyloid beta (A beta), was investigated immunohistochemically. Antibodies to HB-GAM, a cytokine which plays an important role in brain development and maturation, showed strong immunoreactivity with senile plaques in both AD and DS. Anti-HB-GAM reacted with pre-amyloid lesions, but only when markers of dystrophic neurites were present. The presence of HB-GAM in AD brains, but not in control brains, was confirmed by Western blotting. We suggest that the presence of HB-GAM in A beta lesions is a marker of neuronal injury. AD - Department of Pathology, New York University Medical Center, NY 10016, USA. FAU - Wisniewski, T AU - Wisniewski T FAU - Lalowski, M AU - Lalowski M FAU - Baumann, M AU - Baumann M FAU - Rauvala, H AU - Rauvala H FAU - Raulo, E AU - Raulo E FAU - Nolo, R AU - Nolo R FAU - Frangione, B AU - Frangione B LA - eng ID - AG00542/AG/NIA ID - AG05891/AG/NIA ID - NS30455/NS/NINDS ID - etc. PT - Journal Article CY - ENGLAND TA - Neuroreport JID - 9100935 RN - 0 (Amyloid beta-Protein) RN - 0 (Carrier Proteins) RN - 0 (Cytokines) RN - 134034-50-7 (pleiotrophin) SB - IM MH - Adolescent MH - Adult MH - Aged MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein/metabolism MH - Blotting, Western MH - Carrier Proteins/*metabolism MH - Cytokines/*metabolism MH - Down Syndrome/*metabolism MH - Human MH - Immunohistochemistry MH - Middle Age MH - Support, U.S. Gov't, P.H.S. EDAT- 1996/01/31 MHDA- 1996/01/31 00:01 PST - ppublish SO - Neuroreport 1996 Jan 31;7(2):667-71. UI - 96033704 PMID- 7595508 DA - 19951204 DCOM- 19951204 LR - 20001218 IS - 0022-3042 VI - 65 IP - 5 DP - 1995 Nov TI - Affinity purification of proteoglycans that bind to the amyloid protein precursor of Alzheimer's disease. PG - 2201-8 AB - The binding of the amyloid protein precursor (APP) to heparan sulfate proteoglycans has been shown to stimulate the neurite-promoting activity of APP. In this study, proteoglycans that bind with high affinity to APP were characterized. Conditioned medium from cultures of postnatal day 3 mouse brain cells was applied to an affinity column containing a peptide homologous to a heparin-binding domain of APP. A fraction 17-fold enriched in proteoglycans was recovered by elution with a salt gradient. APP bound saturably and with high affinity to the affinity-purified proteoglycan fraction. Scatchard analysis of the binding showed that APP bound to high- and low-affinity sites with equilibrium dissociation constants of 1.4 x 10(-11) and 6.5 x 10(-10) M, respectively. APP, in conjunction with the affinity-purified proteoglycan fraction, promoted neurite outgrowth. The affinity-purified proteoglycan fraction contained a heparan sulfate proteoglycan and a chondroitin sulfate proteoglycan. Digestion of the affinity-purified fraction with heparitinase I revealed a core protein of 63-69-kDa molecular mass, whereas digestion with chondroitinase ABC revealed a core protein of 100-110 kDa. The results suggest that expression of specific APP-binding proteoglycans may be an important step in the regulation of the neurite outgrowth-promoting activity of APP. AD - Department of Pathology, University of Melbourne, Parkville, Victoria, Australia. FAU - Williamson, T G AU - Williamson TG FAU - Nurcombe, V AU - Nurcombe V FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL FAU - Small, D H AU - Small DH LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Proteoglycans) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein Precursor/*metabolism/pharmacology MH - Animal MH - Chromatography, Affinity MH - Mice MH - Neurites/drug effects/physiology MH - Proteoglycans/*isolation & purification/*metabolism/pharmacology MH - Support, Non-U.S. Gov't EDAT- 1995/11/01 MHDA- 1995/11/01 00:01 PST - ppublish SO - J Neurochem 1995 Nov;65(5):2201-8. UI - 96032547 PMID- 7556645 DA - 19951106 DCOM- 19951106 LR - 20021213 IS - 0014-5793 VI - 372 IP - 1 DP - 1995 Sep 18 TI - Glycogen synthase kinase 3 phosphorylates recombinant human tau protein at serine-262 in the presence of heparin (or tubulin). PG - 65-8 AB - Tau protein, the major component of the aberrant structures termed paired helical filaments (PHFs) present in the brain of Alzheimer's disease patients, is pathologically phosphorylated in sites in and around the tubulin-binding sites. A single protein kinase, glycogen synthase kinase 3 (GSK 3), is able to phosphorylate tau at the flanking regions and, additionally, at the tubulin-binding motifs if heparin or tubulin is present. Serines-262 and -324 have been found to be modified at the tubulin-binding region of tau protein by GSK 3 in the presence of heparin or tubulin. AD - Centro de Biologia Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain. FAU - Moreno, F J AU - Moreno FJ FAU - Medina, M AU - Medina M FAU - Perez, M AU - Perez M FAU - Montejo de Garcini, E AU - Montejo de Garcini E FAU - Avila, J AU - Avila J LA - eng PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Peptide Fragments) RN - 0 (Phosphates) RN - 0 (Recombinant Proteins) RN - 0 (Tubulin) RN - 0 (tau Proteins) RN - 56-45-1 (Serine) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Glycogen Synthase Kinase 3) RN - EC 2.7.1.37 (Glycogen Synthase Kinases) SB - IM MH - Amino Acid Sequence MH - Animal MH - Base Sequence MH - Binding Sites MH - Brain/*enzymology MH - Ca(2+)-Calmodulin Dependent Protein Kinase/*metabolism MH - Cattle MH - Glycogen Synthase Kinase 3 MH - Glycogen Synthase Kinases MH - Heparin/*pharmacology MH - Human MH - Microtubules/metabolism MH - Molecular Sequence Data MH - Peptide Fragments/chemistry/metabolism MH - Phosphates/metabolism MH - Phosphorylation MH - Recombinant Proteins/metabolism MH - Serine/metabolism MH - Support, Non-U.S. Gov't MH - Tubulin/metabolism/*pharmacology MH - tau Proteins/chemistry/genetics/*metabolism EDAT- 1995/09/18 MHDA- 1995/09/18 00:01 AID - 0014579395009342 [pii] PST - ppublish SO - FEBS Lett 1995 Sep 18;372(1):65-8. UI - 96159781 PMID- 8588942 DA - 19960327 DCOM- 19960327 LR - 20001218 IS - 0952-3499 VI - 8 IP - 4 DP - 1995 Jul-Aug TI - Characterization of the high affinity heparin binding site of the Alzheimer's disease beta A4 amyloid precursor protein (APP) and its enhancement by zinc(II). PG - 247-57 AB - The Alzheimer's disease beta A4 amyloid precursor protein (APP) has been shown to be involved in a diverse set of biological activities including regulation of cell growth, neurite outgrowth and adhesiveness. The APP and amyloid protein precursor-like proteins (APLP1 and APLP2) belong to a superfamily of proteins that are probably functionally related. In order to characterize the cell adhesion properties of APP the brain specific isoform APP695 was purified and used to assess the binding to heparin, a structural and functional analogue of the glycosaminoglycan heparan sulfate. We show that APP binds in a time dependent and saturable manner to heparin. The salt concentration of 620 mM at which APP elutes from heparin Sepharose is greater than physiological. The apparent equilibrium constant for dissociation was determined to be 300 pM for APP binding to heparin Sepharose. A high affinity heparin binding site was identified within a region conserved in rodent and human APP, APLP1 and APLP2. This binding site was located between residues 316-337 of APP695 which is within the carbohydrate domain of APP. We also demonstrate an interaction between this heparin binding site and the zinc(II) binding site which is conserved in all members of the APP superfamily. We show by using an automated surface plasmon resonance biosensor (BIAcore, Pharmacia) that the affinity for heparin is increased two- to four-fold in the presence of micromolar zinc(II). The identification of zinc-enhanced binding of APP to heparan sulfate side chains of proteoglycans offers a molecular link between zinc(II), as a putative environmental toxin for Alzheimer's disease, and aggregation of amyloid beta A4 protein. AD - Center for Molecular Biology Heidelberg (ZMBH), University of Heidelberg, Germany. FAU - Multhaup, G AU - Multhaup G FAU - Mechler, H AU - Mechler H FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article CY - ENGLAND TA - J Mol Recognit JID - 9004580 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Peptide Fragments) RN - 7440-66-6 (Zinc) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/isolation & purification/*metabolism MH - Animal MH - Binding Sites MH - Biosensing Techniques MH - Brain Chemistry MH - Conserved Sequence MH - Heparin/*metabolism MH - Human MH - Molecular Sequence Data MH - Peptide Fragments/chemical synthesis/chemistry/metabolism MH - Protein Binding MH - Rats MH - Rats, Wistar MH - Sequence Homology, Amino Acid MH - Support, Non-U.S. Gov't MH - Zinc/*metabolism EDAT- 1995/07/01 MHDA- 1995/07/01 00:01 PST - ppublish SO - J Mol Recognit 1995 Jul-Aug;8(4):247-57. UI - 95314326 PMID- 7793988 DA - 19950727 DCOM- 19950727 LR - 20001218 IS - 0003-9861 VI - 320 IP - 1 DP - 1995 Jun 20 TI - Differential binding of vascular cell-derived proteoglycans (perlecan, biglycan, decorin, and versican) to the beta-amyloid protein of Alzheimer's disease. PG - 84-95 AB - Previous studies have demonstrated the immunolocalization of perlecan, a specific heparan sulfate proteoglycan, to the beta-amyloid protein (A beta)-containing amyloid deposits within the walls of blood vessels (i.e., congophilic angiopathy) in Alzheimer's disease (AD) brain. In the present investigation, the differential binding of previously characterized endothelial cell (EC)- and smooth muscle cell (SMC)-derived PGs to A beta was examined to determine whether the accumulation of A beta in cerebrovascular amyloid deposits may be due to its interactions with perlecan. Pretreatment of AA amyloidotic splenic and liver tissue sections with synthetic A beta (1-28) produced strong immunoreactivity with A beta antibodies at tissue sites enriched in perlecan which was partially removed by pretreatment with heparitinase, but not by chondroitin ABC lyase. [35S]-Sulfate labeled proteoglycans (PGs) derived from cultured ECs and SMCs bound to affinity columns containing A beta (1-28) or (1-40), with virtually no binding to A beta (40-1) (reverse peptide), beta-amyloid precursor protein (410-429), or bovine serum albumin. Characterization of EC and SMC PGs bound to A beta (1-28) revealed strong binding by perlecan, weak binding by decorin and biglycan, two dermatan sulfate proteoglycans, and lack of binding by versican/PG-M, a large chondroitin sulfate proteoglycan. Binding of 125I-labeled perlecan to A beta (1-28) was strongly inhibited by isolated perlecan and to a lesser extent by heparin, but not by chondroitin-6-sulfate or unsulfated dextran sulfate. Heparitinase treatment decreased, but did not eliminate the binding of 125I-labeled perlecan to A beta (1-28). Scatchard analysis of the interaction of A beta (1-28)- and EC-derived perlecan in solid-phase assays indicated high-affinity (Kd = 8.3 x 10(-11) M) and lower-affinity (Kd = 4.2 x 10(-8) M) binding sites, with approximately 1 mol of perlecan binding 1.8 mol of A beta. A significant decrease in binding of EC-derived perlecan to A beta (1-28) was observed when a sequence within the putative heparin-binding motif of A beta (His13His14Gln15Lys16) was replaced by the uncharged peptide sequence, Gly13Gly14Gln15Gly16, indicating a perlecan binding site on A beta near the postulated alpha-secretase site (at Lys-16). Overall, the results indicate that specific vascular cell-derived PGs differentially interact with A beta, and that the interactions of highest affinity occur between A beta and binding sites on both the core protein and glycosaminoglycan chains of perlecan.(ABSTRACT TRUNCATED AT 400 WORDS) AD - Department of Pathology, University of Washington, Seattle 98195-6480, USA. FAU - Snow, A D AU - Snow AD FAU - Kinsella, M G AU - Kinsella MG FAU - Parks, E AU - Parks E FAU - Sekiguchi, R T AU - Sekiguchi RT FAU - Miller, J D AU - Miller JD FAU - Kimata, K AU - Kimata K FAU - Wight, T N AU - Wight TN LA - eng ID - AG05136/AG/NIA PT - Journal Article CY - UNITED STATES TA - Arch Biochem Biophys JID - 0372430 RN - 0 (Amyloid beta-Protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Peptide Fragments) RN - 0 (Proteochondroitin Sulfates) RN - 0 (Proteoglycans) RN - 0 (amyloid beta protein (1-28)) RN - 0 (amyloid beta-protein (1-40)) RN - 0 (decorin) RN - 123939-84-4 (biglycan) RN - 126968-45-4 (versican) RN - 143972-95-6 (perlecan) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/genetics/*metabolism MH - Amyloid beta-Protein Precursor/genetics/metabolism MH - Animal MH - Binding Sites MH - Blood Vessels/*metabolism MH - Cattle MH - Female MH - Heparitin Sulfate/metabolism MH - Human MH - In Vitro MH - Kinetics MH - Mice MH - Mice, Inbred CBA MH - Molecular Sequence Data MH - Peptide Fragments/genetics/metabolism MH - Proteochondroitin Sulfates/metabolism MH - Proteoglycans/*metabolism MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1995/06/20 MHDA- 1995/06/20 00:01 AID - S0003986185713458 [pii] PST - ppublish SO - Arch Biochem Biophys 1995 Jun 20;320(1):84-95. UI - 96078821 PMID- 7584721 DA - 19951207 DCOM- 19951207 LR - 20001218 IS - 1340-2544 VI - 15 IP - 3 DP - 1995 Jun TI - [Midkine in relation to development and pathological status of the nervous system] PG - 275-9 AB - Midkine (MK) is a novel heparin-binding growth factor whose expression is regulated by retinoic acid. Midkine is structurally unrelated to other growth factors, and together with pleiotrophin (PTN) subsequently cloned forms a new family of growth factors. Midkine enhances neurite outgrowth and survival of several embryonic neurons. Midkine mRNA is strongly expressed in the nervous system during the midgestation period, while the normal adult brain scarcely expresses it. Immunohistochemical studies revealed midkine localization beside radial glial processes along which neurons migrate. Upon experimental brain infarction in the rat, midkine expression in the ischemic region begins as early as 1 day after the operation. Senile plaques of patients with Alzheimer's disease invariability express midkine. These findings indicate that midkine plays significant roles both in the formation and repair of the central nervous system and processes leading to its diseases. AD - Department of Biochemistry, Nagoya University School of Medicine, Japan. FAU - Muramatsu, T AU - Muramatsu T LA - jpn PT - Journal Article PT - Review PT - Review, Tutorial CY - JAPAN TA - Nihon Shinkei Seishin Yakurigaku Zasshi JID - 9509023 RN - 0 (Carrier Proteins) RN - 0 (Nerve Growth Factors) RN - 0 (Nerve Tissue Proteins) RN - 137497-38-2 (midkine) SB - IM MH - Animal MH - Carrier Proteins/*chemistry/metabolism/physiology MH - English Abstract MH - Human MH - Mice MH - Nerve Growth Factors MH - Nerve Tissue Proteins MH - Nervous System/*embryology MH - Neurites/physiology MH - Rats RF - 26 EDAT- 1995/06/01 MHDA- 1995/06/01 00:01 PST - ppublish SO - Nihon Shinkei Seishin Yakurigaku Zasshi 1995 Jun;15(3):275-9. UI - 95296907 PMID- 7778065 DA - 19950710 DCOM- 19950710 LR - 20001218 IS - 0049-3848 VI - 78 IP - 1 DP - 1995 Apr 1 TI - Zinc (II) selectively enhances the inhibition of coagulation factor XIa by protease nexin-2/amyloid beta-protein precursor. PG - 43-53 AB - Protease nexin-2 (PN-2) is the secreted isoform of the Alzheimer's Amyloid beta-Protein Precursor (A beta PP) that contains the Kunitz-type protease inhibitor (KPI) domain. PN-2/A beta PP is a potent inhibitor of coagulation factor XIa (FXIa) and is secreted in large quantities by activated platelets suggesting a normal function in regulating this protease at sites of vascular injury. In the present study, the effect of Zn2+ on the protease inhibitory properties of PN-2/A beta PP was quantitatively investigated. Zn2+ (1 microM to 1 mM) had no effect on the inhibition of trypsin or chymotrypsin by PN-2/A beta PP. In contrast, Zn2+ at concentrations > 1 microM increased the inhibition of FXIa by PN-2/A beta PP. Enhancement of FXIa inhibition was virtually saturated at approximately 100 microM Zn2+ resulting in a final Ki approximately 6.0 x 10(-11) M. Zn2+ had no effect on the inhibition of FXIa by a purified, recombinant KPI domain of PN-2/A beta PP indicating that the native protein is required for the potentiation of FXIa inhibition. Heparin and Zn2+ were found to further augment each other's ability to stimulate the inhibition of FXIa by PN-2/A beta PP. Together, these findings suggest that the interaction of Zn2+ with PN-2/A beta PP may be important for optimal inhibition of FXIa. AD - Department of Microbiology and Molecular Genetics College of Medicine, University of California, Irvine 92717-4025, USA. FAU - Van Nostrand, W E AU - Van Nostrand WE LA - eng ID - AG00538/AG/NIA ID - HL49566/HL/NHLBI PT - Journal Article CY - UNITED STATES TA - Thromb Res JID - 0326377 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Carrier Proteins) RN - 0 (Cations, Divalent) RN - 0 (Recombinant Fusion Proteins) RN - 0 (protease-nexin) RN - 7440-66-6 (Zinc) RN - 9005-49-6 (Heparin) RN - EC 3.4.21.27 (Factor XIa) SB - IM MH - Amyloid beta-Protein Precursor/*drug effects/pharmacology MH - Carrier Proteins/*drug effects/pharmacology MH - Cations, Divalent MH - Cells, Cultured MH - Factor XIa/*antagonists & inhibitors MH - Heparin/pharmacology MH - Human MH - Recombinant Fusion Proteins/pharmacology MH - Support, U.S. Gov't, P.H.S. MH - Zinc/*pharmacology EDAT- 1995/04/01 MHDA- 1995/04/01 00:01 AID - 004938489500033X [pii] PST - ppublish SO - Thromb Res 1995 Apr 1;78(1):43-53. UI - 95342371 PMID- 7542371 DA - 19950822 DCOM- 19950822 LR - 20011102 IS - 0306-4522 VI - 65 IP - 4 DP - 1995 Apr TI - Expression in mouse embryos and in adult mouse brain of three members of the amyloid precursor protein family, of the alpha-2-macroglobulin receptor/low density lipoprotein receptor-related protein and of its ligands apolipoprotein E, lipoprotein lipase, alpha-2-macroglobulin and the 40,000 molecular weight receptor-associated protein. PG - 1009-25 AB - We have analysed by northern blotting and by in situ hybridization the expression patterns of eight different genes during the second half of mouse embryonic development and in adult mouse brain: we compared the messenger RNA levels of amyloid precursor protein and of the two amyloid precursor protein-like proteins 1 and 2 and we have analysed expression of apolipoprotein E and of its main receptor in brain, the alpha-2-macroglobulin/low density lipoprotein receptor-related protein and three other ligands: the proteinase inhibitor alpha-2-macroglobulin, the modifying enzyme lipoprotein lipase and the 44,000 molecular weight heparin binding protein, a ligand of unknown function. During embryogenesis the temporal expression pattern differs considerably for the three members of the amyloid precursor proteins. Total embryo messenger RNA levels of amyloid precursor protein and amyloid precursor protein-like protein 2 increased progressively, while amyloid precursor protein-like protein 1 messenger RNA showed a burst of synthesis between days 10 and 13 post-coitum. Significantly, expression of the alpha-2-macroglobulin/low density lipoprotein receptor-related protein and of its associated protein, the 44,000 molecular weight heparin binding protein, exhibited their most important increase very similar to that of amyloid precursor protein-like protein 1, between days 10 and 13 post-coitum. Apolipoprotein E, lipoprotein lipase and alpha-2-macroglobulin messenger RNA levels in total embryos increased progressively, beginning most pronounced at days 13, 15 and 17, respectively. In mouse embryos, in situ hybridization established amyloid precursor protein, amyloid precursor protein-like protein 2 and alpha-2-macroglobulin/low density lipoprotein receptor-related protein messenger RNA to be expressed in most organs, with the notable exception of the liver, while expression of the other studied proteins was much more restricted. Among adult mouse tissues, the genes investigated were expressed very prominently in brain, except for lipoprotein lipase and for the complete absence of alpha-2-macroglobulin messenger RNA. In adult mouse brain, the cortex and hippocampus exhibited strong signals for most genes analysed. Exceptions are lipoprotein lipase and apolipoprotein E messenger RNAs, and the absent alpha-2-macroglobulin messenger RNA. Several interesting features, similarities as well as differences, between brain tissue sections hybridized with probes for amyloid precursor protein, amyloid precursor protein-like proteins 1 and 2 and between alpha-2-macroglobulin/low density lipoprotein receptor-related protein and heparin binding protein-44 were observed and are described. The results are further discussed in view of the known or anticipated physiological functions of the proteins examined and of their possible role in the etiology of Alzheimer's disease. AD - Center for Human Genetics, Katholieke Universiteit van Leuven, Belgium. FAU - Lorent, K AU - Lorent K FAU - Overbergh, L AU - Overbergh L FAU - Moechars, D AU - Moechars D FAU - De Strooper, B AU - De Strooper B FAU - Van Leuven, F AU - Van Leuven F FAU - Van den Berghe, H AU - Van den Berghe H LA - eng PT - Journal Article CY - ENGLAND TA - Neuroscience JID - 7605074 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Apolipoproteins E) RN - 0 (Carrier Proteins) RN - 0 (Glycoproteins) RN - 0 (LDL-Receptor Related Protein 1) RN - 0 (LDL-Receptor Related Protein-Associated Protein) RN - 0 (Molecular Chaperones) RN - 0 (Receptors, Immunologic) RN - 0 (Receptors, LDL) RN - 0 (alpha-Macroglobulins) RN - 63231-63-0 (RNA) RN - EC 3.1.1.34 (Lipoprotein Lipase) SB - IM MH - Amyloid beta-Protein Precursor/*biosynthesis/genetics MH - Animal MH - Apolipoproteins E/*biosynthesis MH - Blotting, Northern MH - Brain/embryology MH - Brain Chemistry/*physiology MH - Carrier Proteins/*biosynthesis/genetics MH - Female MH - Gene Expression MH - Glycoproteins/*biosynthesis/genetics MH - In Situ Hybridization MH - LDL-Receptor Related Protein 1 MH - LDL-Receptor Related Protein-Associated Protein MH - Lipoprotein Lipase/biosynthesis/genetics MH - Mice MH - Molecular Chaperones/biosynthesis/genetics MH - Molecular Weight MH - Pregnancy MH - RNA/isolation & purification MH - Receptors, Immunologic/*biosynthesis/genetics MH - Receptors, LDL/*biosynthesis/genetics MH - Support, Non-U.S. Gov't MH - alpha-Macroglobulins/*biosynthesis/genetics EDAT- 1995/04/01 MHDA- 1995/04/01 00:01 AID - 030645229400555J [pii] PST - ppublish SO - Neuroscience 1995 Apr;65(4):1009-25. UI - 95305961 PMID- 7786411 DA - 19950727 DCOM- 19950727 LR - 20001218 IS - 0277-8033 VI - 14 IP - 2 DP - 1995 Feb TI - Tau protein kinase I/GSK-3 beta/kinase FA in heparin phosphorylates tau on Ser199, Thr231, Ser235, Ser262, Ser369, and Ser400 sites phosphorylated in Alzheimer disease brain. PG - 95-105 AB - Previously, tau protein kinase I/glycogen synthase kinase-3 beta/kinase FA(TPKI/GSK-3 beta/FA) was identified as a brain microtubule-associated tau kinase possibly involved in the Alzheimer disease-like phosphorylation of tau. In this report, we find that the TPKI/GSK-3 beta/FA can be stimulated to phosphorylate brain tau up to 8.5 mol of phosphates per mol of protein by heparin, a polyanion compound. Tryptic digestion of 32P-labeled tau followed by high-performance liquid chromatography and high-voltage electrophoresis/thin-layer chromatography reveals 12 phosphopeptides. Phosphoamino acid analysis together with sequential manual Edman degradation and peptide sequence analysis further reveals that TPKI/GSK-3 beta/FA after heparin potentiation phosphorylates tau on sites of Ser199, Thr231, Ser235, Ser262, Ser396, and Ser400, which are potential sites abnormally phosphorylated in Alzheimer tau and potent sites responsible for reducing microtubule binding possibly involved in neuronal degeneration. The results provide initial evidence that TPKI/GSK-3 beta/FA after heparin potentiation may represent one of the most potent systems possibly involved in the abnormal phosphorylation of PHF-tau and neuronal degeneration in Alzheimer disease brains. AD - Institute of Life and Biomedical Sciences, National Tsing Hua University, Hsinchu, Taiwan, ROC. FAU - Song, J S AU - Song JS FAU - Yang, S D AU - Yang SD LA - eng PT - Journal Article CY - UNITED STATES TA - J Protein Chem JID - 8217321 RN - 0 (Phosphopeptides) RN - 0 (tau Proteins) RN - 56-45-1 (Serine) RN - 72-19-5 (Threonine) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.135 (tau-protein kinase) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 2.7.1.37 (Protein-Serine-Threonine Kinases) RN - EC 3.4.21.4 (Trypsin) SB - IM MH - Alzheimer Disease/*enzymology MH - Amino Acid Sequence MH - Animal MH - Brain/*enzymology MH - Cattle MH - Heparin/*pharmacology MH - Microtubules/metabolism MH - Molecular Sequence Data MH - Peptide Mapping MH - Phosphopeptides/chemistry/isolation & purification MH - Phosphorylation MH - Protein Kinases/metabolism MH - Protein-Serine-Threonine Kinases/*metabolism MH - Serine/metabolism MH - Support, Non-U.S. Gov't MH - Swine MH - Threonine/metabolism MH - Trypsin/metabolism MH - tau Proteins/chemistry/*metabolism EDAT- 1995/02/01 MHDA- 1995/02/01 00:01 PST - ppublish SO - J Protein Chem 1995 Feb;14(2):95-105. UI - 95073521 PMID- 7982489 DA - 19950103 DCOM- 19950103 LR - 20001218 IS - 0014-5793 VI - 355 IP - 2 DP - 1994 Nov 28 TI - Interaction between the zinc (II) and the heparin binding site of the Alzheimer's disease beta A4 amyloid precursor protein (APP). PG - 151-4 AB - The Alzheimer's disease beta A4 amyloid precursor protein (APP) has been suggested to be involved in regulation of cell growth, neurite outgrowth and adhesiveness through binding to heparin sulfate proteoglycans. In order to unravel the molecular mechanisms underlying those functions in vitro we show that APP binds in a time dependent and saturable manner to the glycosaminoglycan side-chains of proteoglycans but not to chondroitinsulfate. We also demonstrate an interaction between the high affinity heparin binding site within the carbohydrate domain of APP and the zinc(II) binding site of APP. We show that the affinity for heparin is increased two- to four-fold in the presence of micromolar zinc(II). Thus micromolar concentrations of zinc(II) appear to be able to modulate the binding of APP to heparin side-chains of proteoglycans and as shown previously [Science 265 (1994) 1464-1467] to induce the aggregation of soluble amyloid beta A4 protein. AD - Center for Molecular Biology Heidelberg-ZMBH, University Heidelberg, Germany. FAU - Multhaup, G AU - Multhaup G FAU - Bush, A I AU - Bush AI FAU - Pollwein, P AU - Pollwein P FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Amyloid beta-Protein Precursor) RN - 7440-66-6 (Zinc) RN - 9005-49-6 (Heparin) SB - IM MH - Allosteric Site MH - Alzheimer Disease/metabolism MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Binding Sites MH - Brain/metabolism MH - Heparin/*metabolism MH - Human MH - In Vitro MH - Protein Binding MH - Rats MH - Zinc/*metabolism/pharmacology EDAT- 1994/11/28 MHDA- 1994/11/28 00:01 PST - ppublish SO - FEBS Lett 1994 Nov 28;355(2):151-4. UI - 95014513 PMID- 7929392 DA - 19941123 DCOM- 19941123 LR - 20021101 IS - 0021-9258 VI - 269 IP - 43 DP - 1994 Oct 28 TI - The amyloid beta-protein precursor and its mammalian homologues. Evidence for a zinc-modulated heparin-binding superfamily. PG - 26618-21 AB - The Alzheimer beta-amyloid precursor protein (APP) contains an ectodomain zinc binding site that has been reported to modulate the heparin affinity and protease-inhibitory properties of the molecule. This motif, GVEFVCCP, is highly conserved in amyloid precursor-like proteins 1 and 2 (APLP1 and APLP2), as well as in the Drosophila and Caenorhabditis elegans APP-like proteins (APPL and APL-1). To determine whether the function of this domain is preserved in the human APP-like proteins, the effect of zinc in modulating the elution profile of these proteins upon heparin-Sepharose chromatography was studied. Both APLP1 and APLP2 bound heparin-Sepharose and had NaCl elution profiles similar to that of APP. As previously reported for APP, zinc increased the recovery of APLP1 and APLP2 upon heparin-Sepharose chromatography. APP, APLP1, and APLP2 all bind zinc-chelating Sepharose, indicating that the zinc binding motif may be functionally conserved in these proteins. Additionally, APP, APLP1, and APLP2 migrate at higher molecular sizes (approximately 40 kDa) on SDS-polyacrylamide gel electrophoresis than their predicted molecular sizes. We report data that compare the physicochemical properties of APP to its novel APLP homologues and indicate that these molecules behave as a family of zinc-modulated, heparin-binding proteins. AD - Laboratory of Genetics and Aging, Massachusetts General Hospital, Harvard Medical School, Boston 02129. FAU - Bush, A I AU - Bush AI FAU - Pettingell, W H Jr AU - Pettingell WH Jr FAU - de Paradis, M AU - de Paradis M FAU - Tanzi, R E AU - Tanzi RE FAU - Wasco, W AU - Wasco W LA - eng ID - RO1 AG11899-01/AG/NIA ID - RO1 NS30428-03/NS/NINDS PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (APL-1 protein, C elegans) RN - 0 (APPL protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Helminth Proteins) RN - 0 (Nerve Tissue Proteins) RN - 0 (heparin-sepharose) RN - 7440-66-6 (Zinc) RN - 9012-36-6 (Sepharose) SB - IM GS - ALP1 GS - ALP2 MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/genetics/*metabolism MH - Animal MH - Binding Sites MH - Caenorhabditis elegans MH - Chromatography MH - Comparative Study MH - Conserved Sequence MH - Drosophila MH - Electrophoresis, Polyacrylamide Gel MH - Helminth Proteins/*metabolism MH - Human MH - Mice MH - Molecular Sequence Data MH - Multigene Family MH - Nerve Tissue Proteins/*metabolism MH - Sepharose/analogs & derivatives/metabolism MH - Sequence Homology, Amino Acid MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Zinc/*metabolism EDAT- 1994/10/28 MHDA- 1994/10/28 00:01 PST - ppublish SO - J Biol Chem 1994 Oct 28;269(43):26618-21. UI - 95016672 PMID- 7931292 DA - 19941101 DCOM- 19941101 LR - 20001218 IS - 0022-3042 VI - 63 IP - 4 DP - 1994 Oct TI - Protein kinase FA/glycogen synthase kinase-3 alpha after heparin potentiation phosphorylates tau on sites abnormally phosphorylated in Alzheimer's disease brain. PG - 1416-25 AB - Previously, we identified protein kinase FA/glycogen synthase kinase-3 alpha (GSK-3 alpha) as a brain microtubule-associated tau kinase that phosphorylates Ser235 and Ser404 of tau and causes its electrophoretic mobility shift in gels, a unique property characteristic of paired helical filament-associated pathological tau (PHF-tau) in Alzheimer's disease brains. In this study, we found that the activity of kinase FA/GSK-3 alpha towards phosphorylation of brain tau could be stimulated approximately fourfold by heparin. The phosphorylation molar ratio was increased simultaneously up to 9 mol of phosphates/mol of tau, resulting in a reduced mobility of tau with an apparent molecular mass shift to approximately 68 kDa in sodium dodecyl sulfate gels, which is very similar to that observed in Alzheimer-tau. Tryptic digestion of 32P-labelled tau, followed by HPLC and two-dimensional separation on TLC cellulose plates, revealed eight major phosphopeptides. Phosphoamino acid analysis together with sequential manual Edman degradation and protein sequence analysis further revealed that, in addition to Ser235 and Ser404, heparin generated Thr212, Thr231, Ser262, Ser324, and Ser356, the five extra phosphorylation sites in tau. As Ser235, Ser262, Ser324, Ser356, and Ser404 (particularly the site of Ser262) have been identified as five of the most potent sites in tau responsible for reducing microtubule binding possibly involved in neuronal degeneration, and Thr231, Ser235, Ser262, and Ser404 are four of the most well documented sites abnormally phosphorylated in Alzheimer-tau, the results provide initial evidence that protein kinase FA/GSK-3 alpha after heparin potentiation may represent one of the most potent systems possibly involved in the abnormal phosphorylation of PHF-tau in Alzheimer's disease brains. AD - Institute of Biomedical Sciences, National Tsing Hua University, Hsinchu, Taiwan, R.O.C. FAU - Yang, S D AU - Yang SD FAU - Yu, J S AU - Yu JS FAU - Shiah, S G AU - Shiah SG FAU - Huang, J J AU - Huang JJ LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Peptide Fragments) RN - 0 (Phosphopeptides) RN - 0 (tau Proteins) RN - 1114-81-4 (Phosphothreonine) RN - 17885-08-4 (Phosphoserine) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.123 (Ca(2+)-Calmodulin Dependent Protein Kinase) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 3.4.21.4 (Trypsin) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Animal MH - Brain/*metabolism MH - Ca(2+)-Calmodulin Dependent Protein Kinase/isolation & purification/*metabolism MH - Chromatography, Affinity MH - Comparative Study MH - Heparin/*pharmacology MH - Human MH - Molecular Sequence Data MH - Peptide Fragments/chemistry/isolation & purification MH - Phosphopeptides/chemistry/isolation & purification MH - Phosphorylation MH - Phosphoserine/analysis MH - Phosphothreonine/analysis MH - Protein Kinases/metabolism MH - Substrate Specificity MH - Support, Non-U.S. Gov't MH - Swine MH - Trypsin MH - tau Proteins/chemistry/isolation & purification/*metabolism EDAT- 1994/10/01 MHDA- 1994/10/01 00:01 PST - ppublish SO - J Neurochem 1994 Oct;63(4):1416-25. UI - 94364460 PMID- 8082757 DA - 19941010 DCOM- 19941010 LR - 20001218 IS - 0014-5793 VI - 351 IP - 2 DP - 1994 Sep 5 TI - Secretory cleavage site of Alzheimer amyloid precursor protein is heterogeneous in Down's syndrome brain. PG - 165-7 AB - A beta (beta/A4) is the major constituent of brain amyloid in Alzheimer's disease (AD), Down's syndrome (DS) and normal aged persons. This protein is presumably derived by normal proteolysis from a precursor protein (APP). In this study, C-terminal fragments of APP in a Tris/Triton soluble fraction were partially purified from DS brain by heparin-affinity and reverse phase chromatography, and analyzed by N-terminal amino acid sequencing after SDS polyacrylamide gel electrophoresis and Western blotting. We found at least six different C-terminal fragments including those with the entire A beta region. These results suggest that secretory processing of APP is heterogeneous and generates amyloidogenic C-terminal fragments. AD - Department of Molecular Biology, Tokyo Institute of Psychiatry, Japan. FAU - Kametani, F AU - Kametani F FAU - Tanaka, K AU - Tanaka K FAU - Tokuda, T AU - Tokuda T FAU - Ikeda, S AU - Ikeda S LA - eng PT - Journal Article CY - NETHERLANDS TA - FEBS Lett JID - 0155157 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Peptide Fragments) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/*chemistry/metabolism MH - Brain/metabolism MH - *Brain Chemistry MH - Comparative Study MH - *Down Syndrome/metabolism MH - Female MH - Human MH - Middle Age MH - Molecular Sequence Data MH - Peptide Fragments/*chemistry/metabolism MH - Sequence Analysis EDAT- 1994/09/05 MHDA- 1994/09/05 00:01 PST - ppublish SO - FEBS Lett 1994 Sep 5;351(2):165-7. UI - 94308776 PMID- 7913487 DA - 19940812 DCOM- 19940812 LR - 20001218 IS - 0022-3042 VI - 63 IP - 2 DP - 1994 Aug TI - Secreted form of amyloid beta/A4 protein precursor (APP) binds to two distinct APP binding sites on rat B103 neuron-like cells through two different domains, but only one site is involved in neuritotropic activity. PG - 495-500 AB - Recent studies have identified the Alzheimer's disease amyloid beta/A4 protein precursor (APP) as a trophic and/or tropic protein on several types of cells, including fibroblasts, primary culture neurons, PC12 cells, and B103 neuron-like cells. Many trophic proteins bind heparin, and it is believed that the heparin-binding domain is crucial for the trophic activity of these proteins. APP also binds heparin. The current studies were undertaken to examine the hypothesis that the neuritotropic activity of APP requires heparin binding. It was found that APP produced in E. coli bound B103 cells through detergent-extractable molecules. Approximately 50% of the binding sites were heparinase-sensitive, and heparin and heparan sulfate competed for APP binding to these sites. The heparinase-insensitive sites were recognized by a stretch of 17 amino acids of APP (residues 319-335) that contains the neuritotropic activity of APP. A mutant APP with a deletion at this site was capable of binding to the heparinase-sensitive sites, although this molecule was not neuritotropic to B103 neuron-like cells. Therefore, the neuritotropic site and the heparin-binding site are distinct in APP, and the neuritotropic effect of APP is produced through its binding to detergent-extractable and heparinase-insensitive sites. AD - Department of Neurosciences, School of Medicine, University of California-San Diego, La Jolla 92093-0624. FAU - Ninomiya, H AU - Ninomiya H FAU - Roch, J M AU - Roch JM FAU - Jin, L W AU - Jin LW FAU - Saitoh, T AU - Saitoh T LA - eng ID - AG05131/AG/NIA ID - TW04602/TW/FIC PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Receptors, Cell Surface) RN - 0 (Recombinant Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Binding Sites MH - Cell Line MH - Escherichia coli MH - Heparin/metabolism MH - Kinetics MH - Mutagenesis MH - Neurites/physiology MH - Neurons/*metabolism MH - PC12 Cells MH - Rats MH - Receptors, Cell Surface/*metabolism MH - Recombinant Proteins/metabolism MH - Sequence Deletion MH - Support, U.S. Gov't, P.H.S. EDAT- 1994/08/01 MHDA- 1994/08/01 00:01 PST - ppublish SO - J Neurochem 1994 Aug;63(2):495-500. UI - 95002775 PMID- 7522615 DA - 19941122 DCOM- 19941122 LR - 20001218 IS - 0959-4965 VI - 5 IP - 11 DP - 1994 Jun 27 TI - Binding of heparan sulfate glycosaminoglycan to beta-amyloid peptide: inhibition by potentially therapeutic polysulfated compounds. PG - 1389-92 AB - Heparin sulfate proteoglycans are believed to play an important role in amyloidosis as pathologic chaperones. They bind to amyloidogenic proteins and may mediate the deposition and fibrillogenesis of amyloid at specific tissue sites. In the present study, we demonstrate that heparin sulfate glycosaminoglycan and proteoglycan both bind to the beta-amyloid peptide involved in Alzheimer's disease. The interaction of heparan sulfate proteoglycan and glycosaminoglycan can be inhibited by other sulfated compounds such as heparin, dextran sulfate and pentosan polysulfate. These polysaccharides which are currently used clinically, their derivatives or analogs may be effective as therapeutic agents to prevent or slow the progression of amyloidogenesis in Alzheimer's disease or other amyloidogenic disorders. AD - Department of Geriatrics and Adult Development, Mount Sinai Medical Center, New York, NY 10029-6574. FAU - Leveugle, B AU - Leveugle B FAU - Scanameo, A AU - Scanameo A FAU - Ding, W AU - Ding W FAU - Fillit, H AU - Fillit H LA - eng ID - AI PO1 24671/AI/NIAID ID - AI RO1 24876/AI/NIAID PT - Journal Article CY - ENGLAND TA - Neuroreport JID - 9100935 RN - 0 (Amyloid beta-Protein) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Polysaccharides) RN - 0 (Proteoglycans) RN - 0 (Sulfates) RN - 37300-21-3 (Pentosan Sulfuric Polyester) RN - 9005-49-6 (Heparin) RN - 9042-14-2 (Dextran Sulfate) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Amyloid beta-Protein/*metabolism MH - Binding, Competitive MH - Chromatography, Affinity MH - Comparative Study MH - Dextran Sulfate/pharmacology MH - Heparan Sulfate Proteoglycan MH - Heparin/pharmacology MH - Heparitin Sulfate/isolation & purification/*metabolism MH - Neuroblastoma/chemistry MH - Pentosan Sulfuric Polyester/pharmacology MH - Polysaccharides/*pharmacology MH - Protein Binding/drug effects MH - Proteoglycans/isolation & purification/*metabolism MH - Structure-Activity Relationship MH - Sulfates/*pharmacology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1994/06/27 MHDA- 1994/06/27 00:01 PST - ppublish SO - Neuroreport 1994 Jun 27;5(11):1389-92. UI - 94210049 PMID- 8158260 DA - 19940517 DCOM- 19940517 LR - 20001218 IS - 0270-6474 VI - 14 IP - 4 DP - 1994 Apr TI - A heparin-binding domain in the amyloid protein precursor of Alzheimer's disease is involved in the regulation of neurite outgrowth. PG - 2117-27 AB - The amyloid protein precursor (APP) of Alzheimer's disease is synthesized as an integral transmembrane protein that is released from cells in culture following proteolytic cleavage. The function of released APP is not known, although there is evidence that the protein may bind to components of the extracellular matrix (ECM). In the present study, substratum-bound APP stimulated neurite outgrowth in cultures of chick sympathetic and mouse hippocampal neurons. This effect was dependent upon the presence of substratum-bound heparan sulfate proteoglycans (HSPG). The effect of APP on neurite outgrowth was comparable to that of laminin. A 14 K N-terminal fragment of APP was found to bind heparin and a region close to the N-terminus of APP (residues 96-110) identified as a potential heparin-binding domain based on secondary structure predictions and molecular modeling. Mutagenesis of three basic residues (lysine-99, arginine-100, and arginine-102) resulted in a recombinant protein (APPhep) with decreased heparin-binding capacity. A peptide homologous to the heparin-binding domain was synthesized and found to bind strongly to heparin and to inhibit binding of 125I-labeled APP to heparin (IC50 approximately 10(-7) M). The peptide blocked the effect of APP on neurite outgrowth (IC50 approximately 10(-7) M), whereas two other peptides homologous to other domains in APP had no effect. The results indicate that the binding of APP to HSPG in the ECM may stimulate the effects of APP on neurite outgrowth. AD - Department of Pathology, University of Melbourne, Parkville, Victoria, Australia. FAU - Small, D H AU - Small DH FAU - Nurcombe, V AU - Nurcombe V FAU - Reed, G AU - Reed G FAU - Clarris, H AU - Clarris H FAU - Moir, R AU - Moir R FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurosci JID - 8102140 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (DNA Primers) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Proteoglycans) RN - 0 (Recombinant Proteins) RN - 9005-49-6 (Heparin) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/isolation & purification/*metabolism/*pharmacology MH - Animal MH - Animals, Newborn MH - Base Sequence MH - Binding Sites MH - Brain/metabolism MH - Cattle MH - Chick Embryo MH - Chromatography, Affinity MH - DNA Primers MH - Embryo MH - Ganglia, Sympathetic/cytology/*physiology MH - Heparan Sulfate Proteoglycan MH - Heparin/*metabolism MH - Heparitin Sulfate/isolation & purification/metabolism MH - Hippocampus/cytology/*physiology MH - Human MH - Mice MH - Molecular Sequence Data MH - Mutagenesis, Site-Directed MH - Neurites/drug effects/*physiology/ultrastructure MH - Neurons/cytology/drug effects/*physiology MH - Polymerase Chain Reaction MH - Proteoglycans/isolation & purification/metabolism MH - Recombinant Proteins/isolation & purification/metabolism/pharmacology MH - Restriction Mapping MH - Support, Non-U.S. Gov't EDAT- 1994/04/01 MHDA- 1994/04/01 00:01 PST - ppublish SO - J Neurosci 1994 Apr;14(4):2117-27. UI - 95119135 PMID- 7819340 DA - 19950216 DCOM- 19950216 LR - 20001218 IS - 0300-9084 VI - 76 IP - 3-4 DP - 1994 TI - Identification and regulation of the high affinity binding site of the Alzheimer's disease amyloid protein precursor (APP) to glycosaminoglycans. PG - 304-11 AB - The specific binding of the amyloid protein precursor (APP) to glycosaminoglycans (GAG) suggests that APP is a cell adhesion molecule (CAM) and/or substrate adhesion molecule (SAM). In order to characterize this activity of APP in the brain at the molecular level, we have purified and characterized the major APP species from rat brain. The major isoform isolated was sequenced and found to be APP695. In a solid-phase binding assay, the specificity of this brain-specific APP isoform-GAG interaction was analysed. The binding of APP to the glycosaminoglycan heparin was found to be time-dependent and saturable. A strong heparin-binding site within a region conserved in rodent and human APP, APLP1 and APLP2, was identified. Saturable binding to heparin through this binding site was found to occur at nmol concentrations of APP. This putative high-affinity site was then located within a sequence of 22 amino acids in length corresponding to residues 316-337 of APP695. This sequence is encoded by APP exon 9 and the first three codons of exon 10. Since all APP and L-APP isoforms so far described include these exons, the strong heparin binding site is a ubiquitous feature of all APP and L-APP isoforms strongly suggesting that the brain-specific and neuronal, as well as the non-neuronal and peripheral APPs and L-APPs do have CAM- and SAM-like activities. Certain metal ions including zinc (II) have been proposed as risk factors in Alzheimer's disease (AD). Recently we showed that APP binds zinc (II) at higher nmol concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Center for Molecular Biology, University of Heidelberg, Germany. FAU - Multhaup, G AU - Multhaup G LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - FRANCE TA - Biochimie JID - 1264604 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Glycosaminoglycans) RN - 7440-66-6 (Zinc) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/*chemistry/*metabolism MH - Animal MH - Binding Sites MH - Brain/*metabolism MH - Comparative Study MH - Glycosaminoglycans/chemistry/*metabolism MH - Heparin/*metabolism MH - Human MH - Molecular Sequence Data MH - Rats MH - Sequence Homology, Amino Acid MH - Zinc/metabolism/pharmacology RF - 67 EDAT- 1994/01/01 MHDA- 1994/01/01 00:01 PST - ppublish SO - Biochimie 1994;76(3-4):304-11. UI - 94141321 PMID- 8308393 DA - 19940316 DCOM- 19940316 LR - 20001218 IS - 0037-1017 VI - 65 IP - 12 DP - 1993 Dec TI - [Midkine (MK): a retinoic acid-responsive, heparin-binding growth factor in relationship with differentiation, development, cancer and neural function] PG - 1494-504 AD - Department of Biochemistry, Nagoya University School of Medicine. FAU - Muramatsu, T AU - Muramatsu T LA - jpn PT - Journal Article PT - Review PT - Review Literature CY - JAPAN TA - Seikagaku JID - 0413564 RN - 0 (Carrier Proteins) RN - 0 (Nerve Growth Factors) RN - 0 (Nerve Tissue Proteins) RN - 137497-38-2 (midkine) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/etiology MH - Amino Acid Sequence MH - Animal MH - Carrier Proteins/chemistry/genetics/*physiology MH - Cell Division MH - Gene Expression Regulation MH - Heparin/metabolism MH - Human MH - Molecular Sequence Data MH - Neoplasms, Germ Cell and Embryonal/pathology MH - Nerve Growth Factors MH - Nerve Tissue Proteins MH - Protein Binding MH - Support, Non-U.S. Gov't RF - 50 EDAT- 1993/12/01 MHDA- 1993/12/01 00:01 PST - ppublish SO - Seikagaku 1993 Dec;65(12):1494-504. UI - 94065797 PMID- 8245966 DA - 19931223 DCOM- 19931223 LR - 20001218 IS - 0022-3042 VI - 61 IP - 6 DP - 1993 Dec TI - pH-dependent binding of synthetic beta-amyloid peptides to glycosaminoglycans. PG - 2147-54 AB - The senile plaques found within the cerebral cortex and hippocampus of the Alzheimer disease brain contain beta-amyloid peptide (A beta) fibrils that are associated with a variety of macromolecular species, including dermatan sulfate proteoglycan and heparan sulfate proteoglycan. The latter has been shown recently to bind tightly to both amyloid precursor protein and A beta, and this binding has been attributed largely to the interaction of the core protein of heparan sulfate proteoglycan with A beta and its precursor. Here we have examined the ability of synthetic A beta s to bind to and interact with the glycosaminoglycan moieties of proteoglycans. A beta(1-28) associates with heparin, heparan sulfate, dermatan sulfate, and chondroitin sulfate. The interaction of these sulfated polysaccharides with the amyloid peptide results in the formation of large aggregates that are readily sedimented by centrifugation. The ability of both A beta(1-28) and A beta(1-40) to bind glycosaminoglycans is pH-dependent, with increasing interaction as the pH values fall below neutrality and very little binding at pH 8.0. The pH profile of heparin-induced aggregation of A beta(1-28) has a midpoint pH of approximately 6.5, suggesting that one or more histidine residues must be protonated for binding to occur. Analysis of the A beta sequence reveals a consensus heparin-binding domain at residues 12-17, and this motif contains histidines at positions 13 and 14 that may be involved in the interaction with glycosaminoglycans.(ABSTRACT TRUNCATED AT 250 WORDS) AD - Discovery Research Group, Gliatech Inc., Cleveland, Ohio 44122. FAU - Brunden, K R AU - Brunden KR FAU - Richter-Cook, N J AU - Richter-Cook NJ FAU - Chaturvedi, N AU - Chaturvedi N FAU - Frederickson, R C AU - Frederickson RC LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurochem JID - 2985190R RN - 0 (Amyloid beta-Protein) RN - 0 (Glycosaminoglycans) RN - 0 (Oligopeptides) RN - 0 (Peptide Fragments) RN - 0 (amyloid beta protein (1-28)) RN - 0 (amyloid beta protein (13-17)) RN - 9005-49-6 (Heparin) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Amino Acid Sequence MH - Amyloid beta-Protein/chemistry/isolation & purification/*metabolism MH - Animal MH - Cattle MH - Chromatography, Affinity MH - Comparative Study MH - Glycosaminoglycans/chemistry/*metabolism MH - Heparin/chemistry/metabolism MH - Heparitin Sulfate/chemistry/metabolism MH - Hydrogen-Ion Concentration MH - Molecular Sequence Data MH - Oligopeptides/chemical synthesis MH - Peptide Fragments/chemistry/isolation & purification/*metabolism MH - Protein Binding MH - Structure-Activity Relationship MH - Swine EDAT- 1993/12/01 MHDA- 1993/12/01 00:01 PST - ppublish SO - J Neurochem 1993 Dec;61(6):2147-54. UI - 94115844 PMID- 8287280 DA - 19940223 DCOM- 19940223 LR - 20020419 IS - 0006-8993 VI - 629 IP - 1 DP - 1993 Nov 26 TI - Amyloid beta-protein stimulates casein kinase I and casein kinase II activities. PG - 47-52 AB - Amyloid beta-protein (A beta) is the major protein of cerebrovascular and plaque amyloid in Alzheimer's disease (AD). Extensive evidence has demonstrated abnormal protein phosphorylation in this disease. We investigated the effect of synthetic A beta with the amino-acid sequence corresponding to cerebrovascular A beta and plaque A beta on the activities of casein kinase I (CK I) and casein kinase II (CK II). These enzymes were purified from bovine brain and casein was used as a substrate. A beta was found to stimulate markedly CK I- and CK II-mediated phosphorylation of casein in a concentration-dependent manner. The effect of plaque A beta was considerably higher than that of cerebrovascular A beta. Heparin, which is known to be a specific inhibitor of CK II, completely inhibited A beta-stimulated CK II activity. A beta itself was not a substrate for casein kinases. These findings were confirmed using other substrates for CK I and CK II. The experiments with synthetic CK II-substrate peptide (Leu-Glu-Leu-Ser-Asp-Asp-Asp-Asp-Glu) and the phosphorylation of erythrocyte membrane proteins by intrinsic membrane-bound CK I in erythrocytes showed marked stimulation in activities of casein kinases in the presence of A beta 1-40 or blocked A beta. We propose that A beta, by stimulating casein kinases, may contribute to abnormal protein phosphorylation in AD, in particular to increased phosphorylation of microtubule-associated proteins, leading to the neurofibrillary tangles formation and neurodegeneration in this disease. Interaction of A beta with protein kinases, thus, may characterize the beginning of the disease. AD - Department of Neurochemistry, New York State Institute for Basic Research in Developmental Disabilities, Staten Island 10314-6399. FAU - Chauhan, A AU - Chauhan A FAU - Chauhan, V P AU - Chauhan VP FAU - Murakami, N AU - Murakami N FAU - Brockerhoff, H AU - Brockerhoff H FAU - Wisniewski, H M AU - Wisniewski HM LA - eng ID - GM 47278/GM/NIGMS ID - PO-1-AG04220/AG/NIA PT - Journal Article CY - NETHERLANDS TA - Brain Res JID - 0045503 RN - 0 (Amyloid beta-Protein) RN - 0 (Isoenzymes) RN - 0 (Oligopeptides) RN - EC 2.7.1.- (casein kinase) RN - EC 2.7.1.- (casein kinase II) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 2.7.1.37 (Protein-Serine-Threonine Kinases) SB - IM MH - Amino Acid Sequence MH - Amyloid beta-Protein/*pharmacology MH - Animal MH - Brain/enzymology MH - Cattle MH - Isoenzymes/drug effects/metabolism MH - Kinetics MH - Molecular Sequence Data MH - Oligopeptides/*metabolism MH - Phosphorylation MH - Protein Kinases/drug effects/*metabolism MH - Protein-Serine-Threonine Kinases/drug effects/*metabolism MH - Substrate Specificity MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1993/11/26 MHDA- 1993/11/26 00:01 PST - ppublish SO - Brain Res 1993 Nov 26;629(1):47-52. UI - 94057822 PMID- 8239278 DA - 19931210 DCOM- 19931210 LR - 20001218 IS - 0077-8923 VI - 695 DP - 1993 Sep 24 TI - The role of extracellular matrix in the processing of the amyloid protein precursor of Alzheimer's disease. PG - 169-74 AB - Alzheimer's disease (AD) is characterized by the presence of extracellular amyloid plaques, which contain a protein referred to as the amyloid or beta A4 protein. The beta A4 protein is derived from a larger precursor protein (APP). Studies of autosomal-dominant forms of AD have established the central role of APP in the pathogenesis of the disease. Despite considerable research, the function of APP is unknown. APP can be processed by at least two separate routes. The first route involves a protease known as "APP secretase," which cleaves within the amyloid sequence, thereby mitigating amyloid formation. The second route may result in the production of potentially amyloidogenic fragments. Our studies suggest that following release from the cell membrane, APP interacts with components of the extracellular matrix (ECM) such as the heparan sulfate proteoglycans (HSPG's). The interaction of APP with HSPG's may be important for the function of APP. Substratum-bound APP was found to dramatically increase neurite outgrowth and survival of chick sympathetic neurons in vitro. This effect was dependent upon the presence of substratum-bound HSPG. The results suggest that normally, when bound to the ECM, APP functions to promote neurite outgrowth and/or cell survival. Loss of this normal trophic function might occur in AD, when APP is proteolytically processed via the amyloidogenic pathway. AD - Department of Pathology, University of Melbourne, Victoria, Australia. FAU - Small, D H AU - Small DH FAU - Nurcombe, V AU - Nurcombe V FAU - Clarris, H AU - Clarris H FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Ann N Y Acad Sci JID - 7506858 RN - 0 (Amyloid beta-Protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein/*biosynthesis MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Binding Sites MH - Brain/*metabolism MH - Consensus Sequence MH - Extracellular Matrix/metabolism MH - Heparin/metabolism MH - Human MH - Molecular Sequence Data MH - *Protein Processing, Post-Translational MH - Support, Non-U.S. Gov't RF - 18 EDAT- 1993/09/24 MHDA- 1993/09/24 00:01 PST - ppublish SO - Ann N Y Acad Sci 1993 Sep 24;695:169-74. UI - 93346344 PMID- 8344894 DA - 19930907 DCOM- 19930907 LR - 20001218 IS - 0021-9258 VI - 268 IP - 22 DP - 1993 Aug 5 TI - A novel zinc(II) binding site modulates the function of the beta A4 amyloid protein precursor of Alzheimer's disease. PG - 16109-12 AB - Abnormalities of zinc metabolism occur in Alzheimer's disease (AD), a condition where pathological catabolism of the amyloid protein precursor (APP) causes cerebral beta A4 amyloidosis. An association between zinc and APP metabolism was sought by studying the binding of 65Zn2+ to APP. 65Zn2+ bound in a rapid, saturable, and specific manner (KD = 764 nM). A novel zinc binding motif, strongly conserved between members of the APP family, was located between the cysteine-rich and negatively charged domains of the protein. Zinc increased binding of APP to heparin and has been shown to potentiate the inhibition of coagulation factor XIa by an APP isoform containing a Kunitz-type inhibitory domain (Komiyama, Y., Murakami, T., Egawa, H., Okubo, S., Yasunaga, K., and Murata, K. (1992) Thromb. Res. 66, 397-408) situated near the zinc binding region. Zinc is a factor that modulates the functional properties of the substrate for beta A4 amyloidogenesis. AD - Department of Pathology, University of Melbourne, Parkville, Victoria, Australia. FAU - Bush, A I AU - Bush AI FAU - Multhaup, G AU - Multhaup G FAU - Moir, R D AU - Moir RD FAU - Williamson, T G AU - Williamson TG FAU - Small, D H AU - Small DH FAU - Rumble, B AU - Rumble B FAU - Pollwein, P AU - Pollwein P FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid beta-Protein Precursor) RN - 7440-66-6 (Zinc) SB - IM MH - Alzheimer Disease/*metabolism MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/*metabolism MH - Binding Sites MH - Human MH - Molecular Sequence Data MH - Sequence Homology, Amino Acid MH - Support, Non-U.S. Gov't MH - Zinc/*metabolism EDAT- 1993/08/05 MHDA- 1993/08/05 00:01 PST - ppublish SO - J Biol Chem 1993 Aug 5;268(22):16109-12. UI - 93213269 PMID- 8460999 DA - 19930423 DCOM- 19930423 LR - 20001218 IS - 0006-291X VI - 191 IP - 2 DP - 1993 Mar 15 TI - Secretory form of Alzheimer amyloid precursor protein 695 in human brain lacks beta/A4 amyloid immunoreactivity. PG - 392-8 AB - It is not clear how Alzheimer amyloid precursor proteins (APP) are metabolized in the brain itself. Secretory forms of APP in a phosphate buffer-soluble fraction were purified from post-mortem human brain by heparin-affinity and ion-exchange chromatography and analyzed by N-terminal amino acid sequencing and SDS polyacrylamide gel electrophoresis/immunoblotting. We found apparently similar multi-isoforms of secretory APP (at 93-97, 105-112 and 123 KDa) to those that we have described recently in cerebrospinal fluid. Antisera to the initial part of the beta/A4 sequence labelled only those bands that were found to react with antiserum to the Kunitz-type inhibitor insert of APP, suggesting that beta/A4 amyloid may be generated specifically from APP-695. AD - Department of Molecular Biology, Tokyo Institute of Psychiatry, Japan. FAU - Kametani, F AU - Kametani F FAU - Tanaka, K AU - Tanaka K FAU - Ishii, T AU - Ishii T FAU - Ikeda, S AU - Ikeda S FAU - Kennedy, H E AU - Kennedy HE FAU - Allsop, D AU - Allsop D LA - eng PT - Journal Article CY - UNITED STATES TA - Biochem Biophys Res Commun JID - 0372516 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) SB - IM MH - Adult MH - Aged MH - Alzheimer Disease/metabolism MH - Amino Acid Sequence MH - Amyloid/*metabolism MH - Amyloid beta-Protein Precursor/chemistry/immunology/*metabolism MH - Brain/*metabolism MH - Down Syndrome/metabolism MH - Female MH - Human MH - Male MH - Middle Age MH - Molecular Sequence Data MH - Support, Non-U.S. Gov't EDAT- 1993/03/15 MHDA- 1993/03/15 00:01 AID - S0006291X83712301 [pii] PST - ppublish SO - Biochem Biophys Res Commun 1993 Mar 15;191(2):392-8. UI - 93161094 PMID- 8431762 DA - 19930317 DCOM- 19930317 LR - 20001218 IS - 0006-8993 VI - 601 IP - 1-2 DP - 1993 Jan 22 TI - Binding of secreted human neuroblastoma proteoglycans to the Alzheimer's amyloid A4 peptide. PG - 154-63 AB - Proteoglycans (PGs) may play a fundamental role in all forms of amyloidosis. In Alzheimer's disease, proteoglycans are found deposited in senile plaques and in neurofibrillary tangles. However, the cellular source of these deposited PGs and their role in amyloidosis in Alzheimer's disease is unknown. Proteoglycans were purified from conditioned medium of human neuroblastoma cells (SKNSH-SY 5Y). Two species of proteoglycans were identified by enzyme susceptibility including a heparan sulfate proteoglycan and a dermatan sulfate proteoglycan. A monoclonal antibody to the protein core of a vascular basement membrane heparan sulfate proteoglycan found in senile plaques in Alzheimer's disease cross-reacted with the proteoglycans secreted by human neuroblastoma cells. Binding between 35SO4-labelled neuroblastoma proteoglycans and the Alzheimer amyloid (A4) peptide was demonstrated by affinity chromatography. Specificity studies demonstrated that binding of human neuroblastoma proteoglycans to the amyloid peptide was specific for a heparan sulfate glycosaminoglycan, with some binding to a dermatan sulfate proteoglycan. Binding to A4 was also demonstrated by a chemically deglycosylated protein core preparation. No significant binding of neuroblastoma proteoglycans was found to two other basic peptides derived from the extracellular domain of the beta-amyloid precursor, demonstrating the specificity of proteoglycan binding to the A4 peptide. Human neuroblastoma proteoglycans may bind to the-Alzheimer amyloid A4 peptide in a region with a heparin binding consensus sequence [VHHQKL] which also contains the cleavage site of the beta-amyloid precursor protein. Neuronal proteoglycans may either regulate the secretion of the amyloid protein precursor or modify the binding of the amyloid protein precursor to other cellular adhesion molecules. Alterations in this binding may be related to the pathogenesis of amyloid deposition in Alzheimer's disease. AD - Department of Geriatrics and Adult Development, Mount Sinai Medical Center, New York, NY 10029-6574. FAU - Buee, L AU - Buee L FAU - Ding, W AU - Ding W FAU - Delacourte, A AU - Delacourte A FAU - Fillit, H AU - Fillit H LA - eng ID - AG05138/AG/NIA PT - Journal Article CY - NETHERLANDS TA - Brain Res JID - 0045503 RN - 0 (Amyloid beta-Protein) RN - 0 (Antibodies, Monoclonal) RN - 0 (Proteoglycans) RN - 0 (Sulfates) RN - 56-40-6 (Glycine) RN - 9005-49-6 (Heparin) RN - EC 4.2.2. (Polysaccharide-Lyases) RN - EC 4.2.2.- (Chondroitin Lyases) RN - EC 4.2.2.8 (heparitinsulfate lyase) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein/immunology/*metabolism MH - Antibodies, Monoclonal/diagnostic use MH - Chondroitin Lyases MH - Chromatography, Affinity MH - Electrophoresis, Agar Gel MH - Glycine/metabolism MH - Heparin/metabolism MH - Human MH - Hydrolysis MH - Immunoblotting MH - Immunohistochemistry MH - Nervous System Neoplasms/*metabolism MH - Neuroblastoma/*metabolism MH - Polysaccharide-Lyases MH - Protein Binding MH - Proteoglycans/immunology/isolation & purification/*metabolism MH - Sulfates/metabolism MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Tumor Cells, Cultured EDAT- 1993/01/22 MHDA- 1993/01/22 00:01 PST - ppublish SO - Brain Res 1993 Jan 22;601(1-2):154-63. UI - 93101257 PMID- 1465181 DA - 19930115 DCOM- 19930115 LR - 20011102 IS - 0306-4522 VI - 51 IP - 1 DP - 1992 Nov TI - Acetylcholinesterase and its association with heparan sulphate proteoglycans in cortical amyloid deposits of Alzheimer's disease. PG - 177-84 AB - Previous studies have used a sensitive histochemical technique to demonstrate acetylcholinesterase and butyrylcholinesterase within the pathological lesions of Alzheimer's disease. In this study, we used this technique to show that acetylcholinesterase localized in either frozen or fixed neocortical tissue sections is removed after treatment with various glycosaminoglycans, heparinases or proteases. Heparan sulphate, heparinase lyase type I and to a lesser degree, heparin and chondroitin sulphate were effective in solubilizing a large part of the cholinesterase activity. At physiological concentrations, the protease papain or trypsin readily removed activity but collagenase or pronase were relatively less effective. Peptide protease inhibitors and divalent metals did not exhibit any clear effect. The specificity of these observations was shown by inhibition of activity with various anticholinesterases including diisofluorophosphate. Our results suggest that acetylcholinesterase is anchored to and may be released from the heparan sulphate glycosaminoglycans shown to be contained in the lesions. We further suggest that the localization of cholinesterases is closely associated with the accumulation of the glycosaminoglycans in amyloid plaques and neurofibrillary tangles. AD - Department of Neurology, Case Western Reserve School of Medicine, Cleveland, OH 44106. FAU - Kalaria, R N AU - Kalaria RN FAU - Kroon, S N AU - Kroon SN FAU - Grahovac, I AU - Grahovac I FAU - Perry, G AU - Perry G LA - eng ID - AG 08012/AG/NIA ID - AG07552/AG/NIA ID - AG09287/AG/NIA ID - etc. PT - Journal Article CY - ENGLAND TA - Neuroscience JID - 7605074 RN - 0 (Cholinesterase Inhibitors) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Proteoglycans) RN - 103107-01-3 (Fibroblast Growth Factor 2) RN - 9050-30-0 (Heparitin Sulfate) RN - EC 2.3.1.6 (Choline O-Acetyltransferase) RN - EC 3.1.1.- (Butyrylcholinesterase) RN - EC 3.1.1.7 (Acetylcholinesterase) SB - IM MH - Acetylcholinesterase/*metabolism MH - Aged MH - Alzheimer Disease/*enzymology/pathology MH - Brain/*enzymology/*pathology MH - Butyrylcholinesterase/*metabolism MH - Cerebral Cortex/enzymology MH - Choline O-Acetyltransferase/metabolism MH - Cholinesterase Inhibitors/pharmacology MH - Female MH - Fibroblast Growth Factor 2/metabolism MH - Heparan Sulfate Proteoglycan MH - Heparitin Sulfate/*analysis/pharmacology MH - Human MH - Kinetics MH - Male MH - Postmortem Changes MH - Proteoglycans/*analysis MH - Reference Values MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1992/11/01 MHDA- 1992/11/01 00:01 PST - ppublish SO - Neuroscience 1992 Nov;51(1):177-84. UI - 93057773 PMID- 1279136 DA - 19921211 DCOM- 19921211 LR - 20011102 IS - 0270-6474 VI - 12 IP - 11 DP - 1992 Nov TI - Association and release of the amyloid protein precursor of Alzheimer's disease from chick brain extracellular matrix. PG - 4143-50 AB - The amyloid protein precursor (APP) of Alzheimer's disease was found to bind saturably (Kd = 60 nM) to embryonic chick brain extracellular matrix (ECM). The binding of APP to ECM was not inhibited by 10 micrograms/ml heparin or heparan sulfate. However, pretreatment of cells with 1 mM 4-methylumbelliferyl-beta-D-xyloside, an inhibitor of proteoglycan biosynthesis, reduced the number of APP binding sites on the ECM by 80%. The binding of APP to ECM was also inhibited by pretreatment with chlorate, an inhibitor of glycan sulfation, and heparitinase, which digests the carbohydrate component of heparan sulfate proteoglycans. These results suggest that APP binds with high affinity to one or more heparan sulfate proteoglycans. Acidic and basic fibroblasts growth factor (FGF) also bound to chick ECM. When ECM was incubated with a protease associated with the enzyme AChE (AChE-AP), APP and acidic FGF were released intact from the matrix. The AChE-AP was at least 100-fold more potent in releasing APP from ECM than other trypsin-like proteases (trypsin, plasmin, thrombin). The action of the AChE-AP was inhibited by glia-derived nexin (protease nexin I) and by human brain APP at low nanomolar concentrations. These results suggest that in vivo an AChE-AP may cleave ECM proteins to regulate the availability of soluble APP or other factors bound to the ECM. AD - Department of Pathology, University of Melbourne, Parkville, Victoria, Australia. FAU - Small, D H AU - Small DH FAU - Nurcombe, V AU - Nurcombe V FAU - Moir, R AU - Moir R FAU - Michaelson, S AU - Michaelson S FAU - Monard, D AU - Monard D FAU - Beyreuther, K AU - Beyreuther K FAU - Masters, C L AU - Masters CL LA - eng PT - Journal Article CY - UNITED STATES TA - J Neurosci JID - 8102140 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Protease Inhibitors) RN - 103107-01-3 (Fibroblast Growth Factor 2) RN - 104781-85-3 (Fibroblast Growth Factor 1) RN - EC 3.1.1.8 (Cholinesterases) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein Precursor/antagonists & inhibitors/*metabolism MH - Animal MH - Brain/embryology/*metabolism MH - Chick Embryo/metabolism MH - Cholinesterases/metabolism MH - Extracellular Matrix/*metabolism MH - Fibroblast Growth Factor 1/metabolism MH - Fibroblast Growth Factor 2/metabolism MH - Protease Inhibitors/pharmacology MH - Support, Non-U.S. Gov't EDAT- 1992/11/01 MHDA- 1992/11/01 00:01 PST - ppublish SO - J Neurosci 1992 Nov;12(11):4143-50. UI - 93069537 PMID- 1279864 DA - 19921214 DCOM- 19921214 LR - 20001218 IS - 0166-2236 VI - 15 IP - 10 DP - 1992 Oct TI - Sulfated glycoprotein 2: new relationships of this multifunctional protein to neurodegeneration. PG - 391-6 AB - Sulfated glycoprotein 2 (SGP-2) from rat, and similar molecules from cow, dog, human, pig, ram and quail are known by 11 or more acronyms. SGP-2 is associated with the responses of brain and other tissues to injury; it and related molecules are also normally secreted by the adrenal gland, the liver and the testes. The mRNA of this protein is found in increased levels in Alzheimer's disease. In rats, after perforant path or excitotoxin lesions, levels of the protein or mRNA are elevated in astrocytes, and also in neurons. In rats, brain SGP-2 is regulated by gonadal and adrenal steroids. However, these increases after brain lesions may relate to a function that is associated with the human protein, namely that of inhibiting complement-mediated cell lysis. Other activities suggested for SGP-2 are lipid transport and cell-cell interactions, which are consistent with sequence data that predict binding of dinucleotides, heparin and lipids. The emerging neurobiology of SGP-2 encompasses the subjects of cell death, synaptic remodelling, neuroendocrinology and neurodegenerative diseases. AD - Lilly Research Laboratories, Eli Lilly and Co., Indianapolis, IN 46285. FAU - May, P C AU - May PC FAU - Finch, C E AU - Finch CE LA - eng ID - AG05142/AG/NIA PT - Journal Article PT - Review PT - Review, Tutorial CY - ENGLAND TA - Trends Neurosci JID - 7808616 RN - 0 (Glycoproteins) RN - 0 (clusterin) SB - IM MH - Animal MH - Glycoproteins/*physiology MH - Human MH - Nerve Degeneration/*physiology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. RF - 73 EDAT- 1992/10/01 MHDA- 1992/10/01 00:01 PST - ppublish SO - Trends Neurosci 1992 Oct;15(10):391-6. UI - 92406931 PMID- 1527090 DA - 19921022 DCOM- 19921022 LR - 20001218 IS - 0021-9258 VI - 267 IP - 27 DP - 1992 Sep 25 TI - Phosphorylation of tau protein by purified p34cdc28 and a related protein kinase from neurofilaments. PG - 19705-9 AB - It has been suggested that hyperphosphorylation of the tau protein in neurofibrillary tangles may be relevant to the etiology of Alzheimer's disease and that at least one of the hyperphosphorylated sites lies within a consensus sequence for the p34cdc2/cdc28 family of kinases. We describe a new method for large-scale purification of p34cdc28 kinase from Saccharomyces cerevisiae and show that the purified enzyme can phosphorylate bovine and human tau. Phosphorylation was greatly enhanced by the addition of basic and acidic substrate modulators. The effect of the substrate modulators differed both with the structures of the substrates and the modulators. Similar results were obtained with a kinase that could be purified from neurofilaments by p13suc1 affinity chromatography, a hallmark of p34cdc2/cdc28-type kinases. These results are consistent with the hypothesis that a kinase of this type is involved in tau phosphorylation in vivo and open the possibility that hyperphosphorylation in Alzheimer's disease may be controlled by substrate modulators. AD - Section of Biochemistry and Molecular and Cell Biology, Cornell University, Ithaca, New York 14853. FAU - Mawal-Dewan, M AU - Mawal-Dewan M FAU - Sen, P C AU - Sen PC FAU - Abdel-Ghany, M AU - Abdel-Ghany M FAU - Shalloway, D AU - Shalloway D FAU - Racker, E AU - Racker E LA - eng ID - CA 08964/CA/NCI PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Peptides) RN - 0 (Polysaccharides) RN - 0 (tau Proteins) RN - 123-78-4 (Sphingosine) RN - 25104-18-1 (Polylysine) RN - 25212-18-4 (polyarginine) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.- (p34cdc28 kinase) RN - EC 2.7.1.37 (Protein Kinases) RN - EC 2.7.1.37 (Protein p34cdc2) SB - IM MH - Animal MH - Cattle MH - Heparin/pharmacology MH - Human MH - Intermediate Filaments/*enzymology MH - Kinetics MH - Peptides/pharmacology MH - Phosphorylation/drug effects MH - Polylysine/pharmacology MH - Polysaccharides/pharmacology MH - Protein Kinases/*metabolism MH - Protein p34cdc2/metabolism MH - Saccharomyces cerevisiae/enzymology MH - Sphingosine/pharmacology MH - Spinal Cord/enzymology MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - tau Proteins/*metabolism EDAT- 1992/09/25 MHDA- 1992/09/25 00:01 PST - ppublish SO - J Biol Chem 1992 Sep 25;267(27):19705-9. UI - 92359994 PMID- 1497647 DA - 19920904 DCOM- 19920904 LR - 20001218 IS - 0006-291X VI - 186 IP - 2 DP - 1992 Jul 31 TI - High level expression, purification, and characterization of the Kunitz-type protease inhibitor domain of protease nexin-2/amyloid beta-protein precursor. PG - 1138-45 AB - The protease inhibitor, protease nexin-2 (PN-2), is the secreted isoform of the Alzheimer's amyloid beta-protein precursor (A beta PP) that contains the Kunitz-type protease inhibitor (KPI) domain. Here we describe the use of the methylotrophic industrial yeast Pichia pastoris as a host system for the large scale production of the KPI domain of PN-2/A beta PP. In addition to the 57 amino acid KPI domain, the expression product contained an additional four amino acid residues at its amino terminus that correspond to amino acids 285-288 of A beta PP (Ponte et al. 1988 Nature 311:525-527). This expression system generated yields of greater than 1.0 gram of KPI domain per liter of fermentation media. The secreted 61 amino acid product was purified to homogeneity and biochemically characterized. Amino acid analysis and sequencing of the entire expressed KPI domain verified its integrity. Similar to native PN-2/A beta PP, the purified KPI domain potently inhibited trypsin, chymotrypsin, and coagulation factor XIa. Although heparin augments the inhibition of factor XIa by native PN-2/A beta PP it had no effect on the inhibition of factor XIa by expressed KPI domain suggesting that heparin binds to regions on native PN-2/A beta PP outside of the protease inhibitory domain. This KPI domain expression product should be useful in studying the physiologic and pathophysiologic functions of PN-2/A beta PP. AD - Salk Institute Biotechnology/Industrial Associates, La Jolla, CA 92037-4641. FAU - Wagner, S L AU - Wagner SL FAU - Siegel, R S AU - Siegel RS FAU - Vedvick, T S AU - Vedvick TS FAU - Raschke, W C AU - Raschke WC FAU - Van Nostrand, W E AU - Van Nostrand WE LA - eng ID - AG00538/AG/NIA PT - Journal Article CY - UNITED STATES TA - Biochem Biophys Res Commun JID - 0372516 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Carrier Proteins) RN - 0 (Plasmids) RN - 0 (Recombinant Proteins) RN - 0 (protease-nexin) RN - 9088-41-9 (Trypsin Inhibitor, Kunitz Soybean) RN - EC 3.4.- (Endopeptidases) SB - IM MH - Amino Acid Sequence MH - Amyloid beta-Protein Precursor/*genetics/isolation & purification/pharmacology MH - Base Sequence MH - Carrier Proteins/*genetics/isolation & purification/pharmacology MH - Cells, Cultured MH - Cloning, Molecular MH - Endopeptidases/*metabolism MH - Human MH - Kinetics MH - Molecular Sequence Data MH - Pichia/genetics MH - Plasmids MH - Recombinant Proteins/isolation & purification/pharmacology MH - Restriction Mapping MH - Support, U.S. Gov't, P.H.S. MH - Trypsin Inhibitor, Kunitz Soybean/*genetics/pharmacology EDAT- 1992/07/31 MHDA- 1992/07/31 00:01 PST - ppublish SO - Biochem Biophys Res Commun 1992 Jul 31;186(2):1138-45. UI - 92352227 PMID- 1642473 DA - 19920901 DCOM- 19920901 LR - 20001218 IS - 0364-5134 VI - 32 IP - 1 DP - 1992 Jul TI - An abnormality of plasma amyloid protein precursor in Alzheimer's disease. PG - 57-65 AB - beta A4 amyloid deposition in the brain, which is characteristic of Alzheimer's disease (AD), may result from either overexpression of the amyloid protein precursor (APP) or failure of APP to be correctly processed. A blood marker reflecting this abnormal metabolism would be of diagnostic value and would provide a means of monitoring the efficacy of therapeutic interventions. We analyzed immunoblots of plasma APP enriched by heparin-Sepharose chromatography from patients with moderate to severe AD dementia (n = 34) and control subjects (n = 77) and found an approximately 50% increase in the proportion of 130-kd APP species in patients with AD (p less than 0.001), no difference in the 110-kd form, a 15 to 30% decrease in the 65-kd form (p less than 0.001), and a 20 to 35% decrease in the proportion of 42-kd APP (p less than 0.001). These species of APP were soluble, lacked the carboxyl terminus, and the 110- and 42-kd species were shown to be consistent with degradation products derived from the 130-kd species. A comparison of levels of 130-kd plasma APP from moderately to severely demented patients with AD and control subjects distinguished the two groups with a specificity of 87.0% and a sensitivity of 79.4%. AD - Department of Pathology, University of Melbourne, Australia. FAU - Bush, A I AU - Bush AI FAU - Whyte, S AU - Whyte S FAU - Thomas, L D AU - Thomas LD FAU - Williamson, T G AU - Williamson TG FAU - Van Tiggelen, C J AU - Van Tiggelen CJ FAU - Currie, J AU - Currie J FAU - Small, D H AU - Small DH FAU - Moir, R D AU - Moir RD FAU - Li, Q X AU - Li QX FAU - Rumble, B AU - Rumble B AU - et al. LA - eng PT - Journal Article CY - UNITED STATES TA - Ann Neurol JID - 7707449 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Antibodies, Monoclonal) RN - EC 3.4.21 (Serine Endopeptidases) SB - IM MH - Alzheimer Disease/*blood MH - Amyloid beta-Protein Precursor/*blood/chemistry/metabolism MH - Antibodies, Monoclonal MH - Blotting, Western MH - Human MH - Middle Age MH - Molecular Weight MH - Serine Endopeptidases/metabolism MH - Support, Non-U.S. Gov't EDAT- 1992/07/01 MHDA- 1992/07/01 00:01 PST - ppublish SO - Ann Neurol 1992 Jul;32(1):57-65. UI - 93075943 PMID- 1446027 DA - 19921224 DCOM- 19921224 LR - 20011119 IS - 0365-9615 VI - 113 IP - 6 DP - 1992 Jun TI - [Cytoplasmic casein kinase 2 and its substrate proteins in the brain in Alzheimer's disease] PG - 602-3 AB - The absence of casein kinase 2 on blots of temporal cortex extracts from Alzheimer's disease patients (ADP) was shown using antiserum to casein kinase 2. Casein kinase 2 activity towards endogenous substrates and casein is 2-5 times less in ADP brain in comparison to normal controls. The fractions of heparin-binding proteins, containing protein substrates for phosphorylation, were isolated from temporal cortex of ADP and normal controls. The total amount of heparin-binding proteins from ADP brains is less than from control brains, and the polypeptide composition of these fractions is much more poop. FAU - Aksenova, M V AU - Aksenova MV FAU - Karaseva, M V AU - Karaseva MV FAU - Burbaeva, G Sh AU - Burbaeva GSh LA - rus PT - Clinical Trial PT - Journal Article TT - Tsitoplazmaticheskaia kazeinkinaza 2 i ee substratnye belki v mozge pri bolezni Al'tsgeimera. CY - RUSSIA TA - Biull Eksp Biol Med JID - 0370627 RN - 0 (Nerve Tissue Proteins) RN - 0 (Peptides) RN - EC 2.7.1.- (casein kinase) RN - EC 2.7.1.37 (Protein Kinases) SB - IM MH - Aged MH - Alzheimer Disease/*enzymology MH - Autoradiography MH - Comparative Study MH - Cytoplasm/*enzymology MH - Electrophoresis MH - English Abstract MH - Human MH - Immunoblotting MH - Middle Age MH - Nerve Tissue Proteins/analysis MH - Peptides/analysis MH - Protein Kinases/*analysis MH - Temporal Lobe/*enzymology EDAT- 1992/06/01 MHDA- 1992/06/01 00:01 PST - ppublish SO - Biull Eksp Biol Med 1992 Jun;113(6):602-3. UI - 92330962 PMID- 1628222 DA - 19920818 DCOM- 19920818 LR - 20011102 IS - 0006-8993 VI - 579 IP - 2 DP - 1992 May 8 TI - Basic fibroblast growth factor binds to filamentous inclusions of neurodegenerative diseases. PG - 350-2 AB - The extracellular matrix protein heparin sulfate proteoglycans (HSPG) has been found in the neurofibrillary pathology of Alzheimer disease. This study was performed to determine if similar proteoglycans might be present in the fibrillary inclusions of other neurodegenerative diseases. Basic fibroblast growth factor (bFGF) binding to heparinase sensitive sites was used as an assay for HSPGs. We found that the inclusions of Pick and Parkinson diseases as well as progressive supranuclear palsy contained heparinase sensitive bFGF binding sites while the inclusions of diffuse Lewy body disease lacked bFGF binding sites. These findings indicate that HSPG's interactions and possible role in the formation of intraneuronal inclusions are not limited to Alzheimer disease. AD - Division of Neuropathology, Case Western Reserve University, Cleveland, OH 44106-4901. FAU - Perry, G AU - Perry G FAU - Richey, P AU - Richey P FAU - Siedlak, S L AU - Siedlak SL FAU - Galloway, P AU - Galloway P FAU - Kawai, M AU - Kawai M FAU - Cras, P AU - Cras P LA - eng ID - AG007552/AG/NIA ID - AG009287/AG/NIA ID - K04-AG00415/AG/NIA PT - Journal Article CY - NETHERLANDS TA - Brain Res JID - 0045503 RN - 0 (Proteoglycans) RN - 0 (heparin proteoglycan) RN - 103107-01-3 (Fibroblast Growth Factor 2) RN - 9005-49-6 (Heparin) SB - IM MH - Aged MH - Aged, 80 and over MH - Dementia/metabolism MH - Fibroblast Growth Factor 2/*metabolism MH - Heparin/analogs & derivatives/metabolism MH - Histocytochemistry MH - Human MH - Lewy Bodies/metabolism MH - Middle Age MH - Nerve Degeneration MH - Nervous System Diseases/*metabolism MH - Parkinson Disease/metabolism MH - Proteoglycans/metabolism MH - Support, U.S. Gov't, P.H.S. EDAT- 1992/05/08 MHDA- 1992/05/08 00:01 PST - ppublish SO - Brain Res 1992 May 8;579(2):350-2. UI - 92212933 PMID- 1557394 DA - 19920506 DCOM- 19920506 LR - 20001218 IS - 0027-8424 VI - 89 IP - 7 DP - 1992 Apr 1 TI - A protein kinase associated with paired helical filaments in Alzheimer disease. PG - 2878-82 AB - We have identified a protein kinase in immunoaffinity-purified preparations of paired helical filaments from brain tissue of individuals with Alzheimer disease. The kinase phosphorylates the filament proteins in vitro in a manner independent of second messenger regulation or of modulation by heparin and polyamines. Physiological concentrations of hemin, an oxidized heme porphyrin, inhibit the kinase and abolish Alz-50 immunoreactivity of the proteins. Since paired helical filaments are composed of hyperphosphorylated proteins, association of a protein kinase with the filaments provides a mechanism for abnormal processing of the proteins in disease. AD - Department of Pathology, Albert Einstein College of Medicine, Bronx, NY 10461. FAU - Vincent, I J AU - Vincent IJ FAU - Davies, P AU - Davies P LA - eng ID - AG06803/AG/NIA ID - MH38623/MH/NIMH PT - Journal Article CY - UNITED STATES TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Antibodies, Monoclonal) RN - 0 (Phosphoproteins) RN - 0 (tau Proteins) RN - 16009-13-5 (Hemin) RN - 56-65-5 (Adenosine Triphosphate) RN - EC 2.7.1.37 (Protein Kinases) SB - IM MH - Adenosine Triphosphate/metabolism MH - Alzheimer Disease/*enzymology MH - Antibodies, Monoclonal MH - Cell Compartmentation MH - Hemin/pharmacology MH - Kinetics MH - Microscopy, Electron MH - Neurofibrillary Tangles/*enzymology MH - Phosphoproteins/metabolism MH - Phosphorylation MH - Protein Kinases/antagonists & inhibitors/*metabolism MH - Substrate Specificity MH - Support, U.S. Gov't, P.H.S. MH - tau Proteins/metabolism EDAT- 1992/04/01 MHDA- 1992/04/01 00:01 PST - ppublish SO - Proc Natl Acad Sci U S A 1992 Apr 1;89(7):2878-82. UI - 92392390 PMID- 1520400 DA - 19921014 DCOM- 19921014 LR - 20001218 IS - 1044-7393 VI - 16 IP - 1-2 DP - 1992 Feb-Apr TI - APP-collagen interaction is mediated by a heparin bridge mechanism. PG - 109-21 AB - The amyloid precursor protein (APP) is a glycoprotein consisting of at least four isoforms derived from a single gene by a process of alternative splicing. The membrane-bound forms of APP have been suggested to have adhesive properties and to mediate neural cell adhesion. Previous studies have demonstrated the ability of Fab' fragments of antibodies to extracellular domains of APP to inhibit neural cell binding to a collagen substrate, suggesting a physiological role for the collagen-binding properties of APP. The binding of APP has been demonstrated to be specific for type IV collagen, and no binding to other extracellular matrix components, including fibronectin and laminin, was detected. The APP-collagen binding appeared to be mediated by a heparin-bridge mechanism, since the binding was abolished by the addition of excess heparan or heparinase. These results were observed by both a homogenate-collagen binding assay and a cell-surface adhesion assay, thus providing further evidence for the adhesion role of APP. They also pose the question of the possible role of the heparin-binding properties of APP in the genesis of the neuritic plaques characteristic of Alzheimer's disease. AD - Department of Pharmacology, University College, Dublin, Ireland. FAU - Breen, K C AU - Breen KC LA - eng PT - Journal Article CY - UNITED STATES TA - Mol Chem Neuropathol JID - 8910358 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Extracellular Matrix Proteins) RN - 0 (Immunoglobulins, Fab) RN - 9005-49-6 (Heparin) RN - 9007-34-5 (Collagen) RN - 9050-30-0 (Heparitin Sulfate) RN - EC 4.2.2. (Polysaccharide-Lyases) RN - EC 4.2.2.7 (Heparin Lyase) SB - IM MH - Alzheimer Disease/metabolism MH - Amyloid beta-Protein Precursor/*metabolism MH - Animal MH - Cell Adhesion MH - Collagen/*metabolism MH - Extracellular Matrix Proteins/metabolism MH - Heparin/*metabolism MH - Heparin Lyase MH - Heparitin Sulfate/*metabolism MH - Immunoglobulins, Fab/metabolism MH - Neuroblastoma/pathology MH - Polysaccharide-Lyases/pharmacology MH - Protein Binding MH - Rats MH - Support, Non-U.S. Gov't MH - Tumor Cells, Cultured/drug effects/pathology EDAT- 1992/02/01 MHDA- 1992/02/01 00:01 PST - ppublish SO - Mol Chem Neuropathol 1992 Feb-Apr;16(1-2):109-21. UI - 93093484 PMID- 1459468 DA - 19930114 DCOM- 19930114 LR - 20001218 IS - 0304-324X VI - 38 Suppl 1 DP - 1992 TI - Alzheimer's disease beta-amyloid precursor protein in rat neural cells in culture. PG - 15-23 AB - Immunochemical studies were performed on Alzheimer's beta-amyloid precursor protein (APP) in rat brain and cultured rat neural cells. Multiple APP subtypes were detected on immunoblots of brain homogenate with several antisera specific for subsequences of APP. In rat neural cell cultures, it was demonstrated that the composition of APP subtypes differed among cell types and subcellular fractions, and that APP subtypes in PC12h cells varied in their heparin binding affinity, suggesting distinct functional roles for different APP subtypes. Compatible with the possible role of APP in cell-matrix interaction, an increase in oligodendroglial APP was observed following their attachment onto poly-L-lysine substratum. AD - Department of Medicine, University of British Columbia, Vancouver, Canada. FAU - Mizuguchi, M AU - Mizuguchi M FAU - Ikeda, K AU - Ikeda K FAU - Namba, Y AU - Namba Y FAU - Kim, S U AU - Kim SU LA - eng PT - Journal Article CY - SWITZERLAND TA - Gerontology JID - 7601655 RN - 0 (Amyloid beta-Protein Precursor) RN - 9005-49-6 (Heparin) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid beta-Protein Precursor/classification/*metabolism MH - Animal MH - Brain/metabolism MH - Cell Adhesion MH - Cells, Cultured MH - Comparative Study MH - Heparin/metabolism MH - Human MH - Neurons/metabolism MH - Oligodendroglia/cytology/metabolism MH - Protein Binding MH - Rats MH - Subcellular Fractions/metabolism MH - Support, Non-U.S. Gov't EDAT- 1992/01/01 MHDA- 1992/01/01 00:01 PST - ppublish SO - Gerontology 1992;38 Suppl 1:15-23. UI - 91302299 PMID- 1906461 DA - 19910819 DCOM- 19910819 LR - 20001218 IS - 0021-9258 VI - 266 IP - 20 DP - 1991 Jul 15 TI - High affinity interactions between the Alzheimer's beta-amyloid precursor proteins and the basement membrane form of heparan sulfate proteoglycan. PG - 12878-83 AB - High affinity interactions were studied between the basement membrane form of heparan sulfate proteoglycan (HSPG) and the 695-, 751-, and 770-amino acid Alzheimer amyloid precursor (AAP) proteins. Based on quantitative analyses of binding data, we identified single binding sites for the HSPG on AAP-695 (Kd = 9 x 10(-10) M), AAP-751 (Kd = 10 x 10(-9) M), and AAP-770 (Kd = 9 x 10(-9) M). It is postulated that the "Kunitz" protease inhibitor domain which is present in AAP-751 and -770 reduces the affinity of AAPs for the HSPG through steric hindrance and/or conformational alteration. HSPG binding was inhibited by heparin and dextran sulfate, but not by dermatan or chondroitin sulfate. HSPG protein core, obtained by heparitinase digestion, also bound to the beta-amyloid precursor proteins with high affinity, indicating that the high affinity binding site is constituted by the polypeptide chain rather than the carbohydrate moiety. The effects of various cations on these interactions were also studied. Our results suggest that specific interactions between the AAP proteins and the extracellular matrix may be involved in the nucleation stages of Alzheimer's disease type amyloidogenesis. AD - Department of Pathology, Queen's University, Syl & Molly Apps Research Center, Kingston General Hospital, Ontario, Canada. FAU - Narindrasorasak, S AU - Narindrasorasak S FAU - Lowery, D AU - Lowery D FAU - Gonzalez-DeWhitt, P AU - Gonzalez-DeWhitt P FAU - Poorman, R A AU - Poorman RA FAU - Greenberg, B AU - Greenberg B FAU - Kisilevsky, R AU - Kisilevsky R LA - eng PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid beta-Protein) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Fibronectins) RN - 0 (Heparan Sulfate Proteoglycan) RN - 0 (Protease Inhibitors) RN - 0 (Protein Precursors) RN - 0 (Proteochondroitin Sulfates) RN - 0 (Recombinant Proteins) RN - 9050-30-0 (Heparitin Sulfate) SB - IM MH - Alzheimer Disease/metabolism MH - Amyloid beta-Protein/genetics/*metabolism MH - Amyloid beta-Protein Precursor MH - Basement Membrane/metabolism MH - Binding Sites MH - Fibronectins/metabolism MH - Heparan Sulfate Proteoglycan MH - Heparitin Sulfate/isolation & purification/*metabolism MH - Human MH - Kinetics MH - Protease Inhibitors/*metabolism MH - Protein Binding MH - Protein Precursors/genetics/*metabolism MH - Proteochondroitin Sulfates/isolation & purification/*metabolism MH - Recombinant Proteins/metabolism MH - Support, Non-U.S. Gov't EDAT- 1991/07/15 MHDA- 1991/07/15 00:01 PST - ppublish SO - J Biol Chem 1991 Jul 15;266(20):12878-83. UI - 91324686 PMID- 1865106 DA - 19910906 DCOM- 19910906 LR - 20011102 IS - 0022-1554 VI - 39 IP - 7 DP - 1991 Jul TI - Basic fibroblast growth factor binding is a marker for extracellular neurofibrillary tangles in Alzheimer disease. PG - 899-904 AB - Neurofibrillary tangles (NFT) are abnormal filamentous inclusions that develop in neurons in Alzheimer disease and other disorders. When neurons die, the neurofibrillary tangles that persist in the extracellular space show ultrastructural and antigenic changes. Both intra- and extracellular NFT have recently been shown to contain heparan sulfate proteoglycans (HSPGs). HSPGs are also present in other amyloid deposits in the brain and in systemic amyloidoses. Basic fibroblast growth factor (bFGF) is a heparin binding growth factor which is involved in angiogenesis and also has neurite promoting activity. We now report that bFGF binds avidly to extracellular NFT. Alz-50, a monoclonal antibody (MAb) to an abnormal form of tau and bFGF binding label mutually exclusive subpopulations of neurofibrillary tangles. bFGF binding is abolished by heparinase or heparitinase treatment and therefore is most likely based on the presence of HSPG. Binding of bFGF is a specific and sensitive morphological method to distinguish intra- from extracellular NFT. As intracellular NFT, which also contain HSPGs, are not labeled by bFGF binding, this finding also suggests that HSPGs are modified when the NFT become extracellular. AD - Division of Neuropathology, Case Western Reserve University, Cleveland, Ohio 44106. FAU - Siedlak, S L AU - Siedlak SL FAU - Cras, P AU - Cras P FAU - Kawai, M AU - Kawai M FAU - Richey, P AU - Richey P FAU - Perry, G AU - Perry G LA - eng ID - AG-007552/AG/NIA ID - K04-AG00415/AG/NIA PT - Journal Article CY - UNITED STATES TA - J Histochem Cytochem JID - 9815334 RN - 0 (Biological Markers) RN - 103107-01-3 (Fibroblast Growth Factor 2) SB - IM MH - Aged MH - Aged, 80 and over MH - Alzheimer Disease/metabolism/*pathology MH - Biological Markers MH - Fibroblast Growth Factor 2/*metabolism MH - Hippocampus/metabolism/*ultrastructure MH - Human MH - Microscopy, Electron MH - Neurofibrils/*metabolism/ultrastructure MH - Support, U.S. Gov't, P.H.S. MH - Temporal Lobe/metabolism/*ultrastructure EDAT- 1991/07/01 MHDA- 1991/07/01 00:01 PST - ppublish SO - J Histochem Cytochem 1991 Jul;39(7):899-904. UI - 92145975 PMID- 1685979 DA - 19920319 DCOM- 19920319 LR - 20001218 IS - 0300-9629 VI - 100 IP - 3 DP - 1991 TI - A heparin-binding protein from neuroblastoma cells: immunological comparison to beta-amyloid precursor protein. PG - 715-8 AB - 1. beta-Amyloid precursor protein cross-reactive polypeptides were detected in the membrane extracts of a mouse neuroblastoma cell line, NB41A3. Four immunoreactive polypeptide bands were observed on western blots of a cell membrane extract. Their molecular weights as estimated by polyacrylamide gel electrophoresis ranged from 89.1 to 41 kDa. 2. After heparin affinity chromatography, two of these polypeptides strongly cross-reacted with an antibody that recognizes Alzheimer beta-amyloid precursor protein. 3. From the heparin binding fraction, these protein were further separated by reverse-phase high-performance liquid chromatography. A cross-reactive protein was isolated. AD - Department of Chemistry, University of Alaska Fairbanks 99775-0180. FAU - Zhao, X H AU - Zhao XH FAU - Schoenheit, C AU - Schoenheit C FAU - Duffy, L K AU - Duffy LK LA - eng ID - R15AG08978/AG/NIA PT - Journal Article CY - ENGLAND TA - Comp Biochem Physiol A JID - 1276312 RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Carrier Proteins) RN - 9005-49-6 (Heparin) SB - IM MH - Amyloid beta-Protein Precursor/*analysis MH - Animal MH - Blotting, Western MH - Carrier Proteins/*analysis MH - Chromatography, Affinity MH - Chromatography, High Pressure Liquid MH - Comparative Study MH - Cross Reactions MH - Heparin/*metabolism MH - Mice MH - Neuroblastoma/chemistry MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, Non-P.H.S. MH - Support, U.S. Gov't, P.H.S. MH - Tumor Cells, Cultured/*chemistry EDAT- 1991/01/01 MHDA- 1991/01/01 00:01 PST - ppublish SO - Comp Biochem Physiol A 1991;100(3):715-8. UI - 90277634 PMID- 2112543 DA - 19900716 DCOM- 19900716 LR - 20001218 IS - 0021-9258 VI - 265 IP - 17 DP - 1990 Jun 15 TI - Immunopurification and protease inhibitory properties of protease nexin-2/amyloid beta-protein precursor. PG - 9591-4 AB - Protease nexin-2 (PN-2) is a protease inhibitor that is synthesized and secreted by a variety of extravascular cells including human fibroblasts. It forms sodium dodecyl sulfate-stable complexes with trypsin, the epidermal growth factor binding protein and the gamma-subunit of nerve growth factor. Recently we reported that PN-2 is the secreted form of the amyloid beta-protein precursor (APP) and is a potent inhibitor of chymotrypsin. Here we describe a two-step procedure to purify PN-2/APP using a monoclonal antibody immunoaffinity column. We also quantitated the protease inhibitory properties of purified PN-2/APP on a number of serine proteases. PN-2/APP was a potent inhibitor of coagulation factor XIa with a Ki = 2.9 x 10(-10). The inhibition of factor XIa by PN-2/APP was augmented by heparin and resulted in a Ki = 5.5 x 10(-11) M. Trypsin and chymotrypsin were also effectively inhibited with a Ki = 4.2 x 10(-10) and 1.6 x 10(-9), respectively. PN-2/APP also inhibited the epidermal growth factor binding protein, the gamma-subunit of nerve growth factor, and chymase and plasmin to a lesser extent. In view of recent findings that PN-2/APP is contained in alpha-granules of platelets and is secreted upon platelet activation, the potent inhibition of factor XIa suggests that PN-2/APP may play a regulatory role in the coagulation pathway at vascular wound sites. In addition, these studies define biochemical activities of PN-2/APP which may be involved in regulating proteases that lead to the generation and deposition of the beta-protein in neurodegenerative lesions associated with Alzheimer's disease and Down's syndrome. AD - Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717. FAU - Van Nostrand, W E AU - Van Nostrand WE FAU - Wagner, S L AU - Wagner SL FAU - Farrow, J S AU - Farrow JS FAU - Cunningham, D D AU - Cunningham DD LA - eng ID - GM-31609/GM/NIGMS PT - Journal Article CY - UNITED STATES TA - J Biol Chem JID - 2985121R RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Carrier Proteins) RN - 0 (Protease Inhibitors) RN - 0 (Protein Precursors) RN - 0 (protease-nexin) RN - EC 3.4 (Peptide Hydrolases) SB - IM MH - Amino Acid Sequence MH - Amyloid/*isolation & purification/pharmacology MH - Amyloid beta-Protein Precursor MH - Carrier Proteins/*isolation & purification/pharmacology MH - Cells, Cultured MH - Chromatography, Affinity MH - Fibroblasts/metabolism MH - Human MH - Infant, Newborn MH - Kinetics MH - Male MH - Molecular Sequence Data MH - Molecular Weight MH - Peptide Hydrolases/metabolism MH - Protease Inhibitors/*isolation & purification MH - Protein Precursors/*isolation & purification/pharmacology MH - Skin/metabolism MH - Substrate Specificity MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. EDAT- 1990/06/15 MHDA- 1990/06/15 00:01 PST - ppublish SO - J Biol Chem 1990 Jun 15;265(17):9591-4. UI - 90260664 PMID- 2111585 DA - 19900626 DCOM- 19900626 LR - 20010323 IS - 0036-8075 VI - 248 IP - 4959 DP - 1990 Jun 1 TI - Platelet coagulation factor XIa-inhibitor, a form of Alzheimer amyloid precursor protein. PG - 1126-8 AB - An inhibitor of coagulation factor XIa was purified from serum-free conditioned medium of HepG2 liver cells. Platelets stimulated with thrombin or calcium ionophore (A23187) secrete a protein functionally and immunologically identical to the inhibitor, implying a role for this inhibitor in hemostasis. Analysis of the amino-terminal amino acid sequence and immunologic reactivity showed the inhibitor to be a truncated form of the Alzheimer's amyloid precursor protein that contains a Kunitz-type serine protease inhibitor domain and at least a portion of the amyloid beta protein. It inhibits factor XIa and trypsin with a Ki of 450 +/- 50 pM and 20 +/- 10 pM, respectively. Heparin (1 unit/ml) did not significantly effect inhibition of trypsin, but inhibition of XIa was 15 times greater (Ki = 25 +/- 15 pM) in the presence of heparin. AD - Department of Medicine, Jewish Hospital, Washington University Medical Center, St. Louis, MO 63110. FAU - Smith, R P AU - Smith RP FAU - Higuchi, D A AU - Higuchi DA FAU - Broze, G J Jr AU - Broze GJ Jr LA - eng PT - Journal Article CY - UNITED STATES TA - Science JID - 0404511 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Blood Proteins) RN - 0 (Protein Precursors) RN - 0 (Serine Proteinase Inhibitors) RN - EC 3.4.21.27 (Factor XIa) SB - IM MH - Amino Acid Sequence MH - Amyloid/immunology MH - Amyloid beta-Protein Precursor MH - Blood Platelets/*analysis MH - Blood Proteins/immunology/*isolation & purification/physiology MH - Cross Reactions MH - Factor XIa/*antagonists & inhibitors MH - Human MH - Molecular Sequence Data MH - Protein Precursors/immunology MH - Serine Proteinase Inhibitors/blood/*isolation & purification EDAT- 1990/06/01 MHDA- 2001/03/28 10:01 PST - ppublish SO - Science 1990 Jun 1;248(4959):1126-8. UI - 89184568 PMID- 2494659 DA - 19890428 DCOM- 19890428 LR - 20001218 IS - 0027-8424 VI - 86 IP - 6 DP - 1989 Mar TI - Characterization of an amyloid beta precursor protein that binds heparin and contains tyrosine sulfate. PG - 2066-9 AB - A secreted form of the amyloid beta protein precursor was isolated from the growth conditioned medium of the PC12 sympathetic nerve-like cell line. This protein is recognized by an antiserum that detects a protein of 140 kDa and a less abundant species of 115 kDa on NaDodSO4/acrylamide gels. The amyloid precursor proteins contain O-linked sugars and tyrosine sulfate and bind to the glycosaminoglycan heparin. These results suggest a role for extracellular sulfated glycoproteins in the pathogenesis of Alzheimer disease. AD - Salk Institute for Biological Studies, San Diego, CA 92138. FAU - Schubert, D AU - Schubert D FAU - LaCorbiere, M AU - LaCorbiere M FAU - Saitoh, T AU - Saitoh T FAU - Cole, G AU - Cole G LA - eng ID - AG 05131/AG/NIA ID - F2 AG 05424A/AG/NIA ID - NS 09658/NS/NINDS PT - Journal Article CY - UNITED STATES TA - Proc Natl Acad Sci U S A JID - 7505876 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Carbohydrates) RN - 0 (Peptide Fragments) RN - 0 (Protease Inhibitors) RN - 0 (Protein Precursors) RN - 55520-40-6 (Tyrosine) RN - 9005-49-6 (Heparin) RN - 956-46-7 (tyrosine O-sulfate) SB - IM MH - Adrenal Gland Neoplasms/metabolism MH - Alzheimer Disease MH - Amyloid/analysis/*metabolism MH - Amyloid beta-Protein Precursor MH - Animal MH - Blotting, Western MH - Carbohydrates/analysis MH - Electrophoresis, Polyacrylamide Gel MH - Heparin/*metabolism MH - Immunoassay MH - Molecular Weight MH - Peptide Fragments/analysis/metabolism MH - Pheochromocytoma/metabolism MH - Protease Inhibitors MH - Protein Precursors/analysis/*metabolism MH - Rats MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. MH - Tumor Cells, Cultured MH - Tyrosine/*analogs & derivatives/analysis EDAT- 1989/03/01 MHDA- 1989/03/01 00:01 PST - ppublish SO - Proc Natl Acad Sci U S A 1989 Mar;86(6):2066-9. UI - 90099536 PMID- 2513585 DA - 19900131 DCOM- 19900131 LR - 20001218 IS - 0361-7742 VI - 317 DP - 1989 TI - Evidence for the derivation of a peptide ligand from the amyloid beta-protein precursor of Alzheimer's disease. PG - 893-902 AB - A monoclonal antibody to a synthetic peptide consisting of residues 8-17 of amyloid beta-protein was used in immunohistochemical studies to reveal binding sites for this peptide in cytoplasmic vesicles in cells of the adrenal zona reticularis and the islets of Langerhans of the pancreas. These binding sites showed some specificity for this peptide and so may represent a membrane receptor. These results suggest that the membrane bound beta-protein precursor may be processed by limited extracellular proteolysis to release a peptide ligand containing the 8-17 sequence. It has been reported recently that the core protein of a heparin sulphate proteoglycan (HSPG) secreted by PC12 cells shows some homology with the beta-protein precursor. This suggests that the binding sites might be due to the presence of a HSPG core protein receptor. Further studies should be carried out to find out if receptors to beta-protein peptides are present in brain tissue, since these might play a role in the catabolism of the beta-protein precursor, and in the formation of cerebral amyloid in Alzheimer's disease (AD). AD - Department of Pathology M-012, University of California, San Diego School of Medicine, La Jolla 92093. FAU - Allsop, D AU - Allsop D FAU - Ikeda, S AU - Ikeda S FAU - Glenner, G G AU - Glenner GG LA - eng ID - AG05683/AG/NIA PT - Journal Article PT - Review PT - Review, Tutorial CY - UNITED STATES TA - Prog Clin Biol Res JID - 7605701 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Protein Precursor) RN - 0 (Ligands) RN - 0 (Protein Precursors) SB - IM MH - Alzheimer Disease/*metabolism MH - Amyloid/*analysis MH - Amyloid beta-Protein Precursor MH - Binding Sites MH - Human MH - Ligands MH - Protein Precursors/*analysis MH - Support, Non-U.S. Gov't MH - Support, U.S. Gov't, P.H.S. RF - 27 EDAT- 1989/01/01 MHDA- 1989/01/01 00:01 PST - ppublish SO - Prog Clin Biol Res 1989;317:893-902. UI - 89238360 PMID- 2469952 DA - 19890619 DCOM- 19890619 LR - 20001218 IS - 0077-0094 VI - 23 DP - 1989 TI - Ateroid in the treatment of dementia: results of a clinical trial. PG - 76-84 AD - University of Pisa, Institute of Clinical Psychiatry, Italy. FAU - Conti, L AU - Conti L FAU - Placidi, G F AU - Placidi GF FAU - Cassano, G B AU - Cassano GB LA - eng PT - Clinical Trial PT - Journal Article CY - SWITZERLAND TA - Mod Probl Pharmacopsychiatry JID - 0425142 RN - 0 (Antilipemic Agents) RN - 0 (Glycosaminoglycans) RN - 0 (Heparinoids) SB - IM MH - Aged MH - Alzheimer Disease/*drug therapy/psychology MH - Antilipemic Agents/*therapeutic use MH - Clinical Trials MH - Dementia, Multi-Infarct/*drug therapy/psychology MH - Female MH - Glycosaminoglycans/*therapeutic use MH - Heparinoids/*therapeutic use MH - Human MH - Injections, Intravenous MH - Male MH - Neuropsychological Tests EDAT- 1989/01/01 MHDA- 1989/01/01 00:01 PST - ppublish SO - Mod Probl Pharmacopsychiatry 1989;23:76-84.