CA2+-DEPENDENT 68-KILODALTON PROTEASE IN FAMILIAL ALZHEIMERS-DISEASE CELLS CLEAVES THE N-TERMINUS OF BETA-AMYLOID

被引:18
作者
MATSUMOTO, A
FUJIWARA, Y
机构
[1] Department of Radiation Biophysics and Genetics, Kobe University School of Medicine, Kobe 650, Kusunoki-cho 7-5-1, Chuo-Ku
关键词
D O I
10.1021/bi00179a021
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Lymphoblastoid cells derived from patients with early- and late-onset familial Alzheimer's disease express a Ca2+-dependent, 68-kDa protease which forms an SDS-stable and heat-labile complex with the beta-amyloid precursor protein. Utilizing this property, we prepared the protein by heat-dissociation of its immunoprecipitate with an antibody raised against the extracellular part of the beta-amyloid precursor protein. Disuccinimidyl suberate cross-linking analysis showed that in the presence of Ca2+ this protein binds to a synthetic oligopeptide corresponding to the first 12 amino acids of beta/A4-amyloid and its N-terminal flanks. Thin-layer chromatography of a reaction mixture of the 68-kDa protein and the oligopeptide demonstrated its proteolytic activity in the presence of Ca2+. Subsequent N-terminal amino acid sequencing of the cleaved fragment showed the cleavage site of the oligopeptide to be the Lys-2-Met-1-Asp(betaA4-1) bonds. This protease also cleaves a natural substrate of 110-kDa beta-amyloid precursor protein, thereby generating the 16-kDa preamyloid peptide that accumulates abnormally in familial Alzheimer's disease lymphoblastoid cells. It does not, however, cleave the Gln(betaA4-15)-Lys16-Leu17 bond that is regarded to be the normal proteolytic site for the secretion of the beta-amyloid precursor protein. Analysis of the effects of protease inhibitors suggests that this 68-kDa protease is a Ca2+-dependent serine protease.
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页码:3941 / 3948
页数:8
相关论文
共 40 条
[1]  
ABRAHAM C R, 1990, Peptide Research, V3, P211
[2]   PURIFICATION AND CLONING OF BRAIN PROTEASES CAPABLE OF DEGRADING THE BETA-AMYLOID PRECURSOR PROTEIN [J].
ABRAHAM, CR ;
RAZZABONI, BL ;
PAPASTOITSIS, G ;
PICARD, E ;
KANEMARU, K ;
MECKELEIN, B ;
MUCKE, L .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1992, 674 :174-179
[3]   A CALCIUM-ACTIVATED PROTEASE FROM ALZHEIMERS-DISEASE BRAIN CLEAVES AT THE N-TERMINUS OF THE AMYLOID BETA-PROTEIN [J].
ABRAHAM, CR ;
DRISCOLL, J ;
POTTER, H ;
VANNOSTRAND, WE ;
TEMPST, P .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 174 (02) :790-796
[4]   ALZHEIMER AMYLOID BETA/A4 PEPTIDE BINDING-SITES AND A POSSIBLE APP-SECRETASE ACTIVITY ASSOCIATED WITH RAT-BRAIN CORTICAL MEMBRANES [J].
ALLSOP, D ;
YAMAMOTO, T ;
KAMETANI, F ;
MIYAZAKI, N ;
ISHII, T .
BRAIN RESEARCH, 1991, 551 (1-2) :1-9
[5]  
BEYREUTHER K, 1991, ANN NY ACAD SCI, V640, P129
[6]  
BIRD TD, 1989, ANN NEUROL, V25, P487
[7]   MUTATION OF THE BETA-AMYLOID PRECURSOR PROTEIN IN FAMILIAL ALZHEIMERS-DISEASE INCREASES BETA-PROTEIN PRODUCTION [J].
CITRON, M ;
OLTERSDORF, T ;
HAASS, C ;
MCCONLOGUE, L ;
HUNG, AY ;
SEUBERT, P ;
VIGOPELFREY, C ;
LIEBERBURG, I ;
SELKOE, DJ .
NATURE, 1992, 360 (6405) :672-674
[8]  
Creighton TE, 1989, PROTEIN STRUCTURE PR, P225
[9]   CLEAVAGE OF AMYLOID-BETA PEPTIDE DURING CONSTITUTIVE PROCESSING OF ITS PRECURSOR [J].
ESCH, FS ;
KEIM, PS ;
BEATTIE, EC ;
BLACHER, RW ;
CULWELL, AR ;
OLTERSDORF, T ;
MCCLURE, D ;
WARD, PJ .
SCIENCE, 1990, 248 (4959) :1122-1124
[10]   POTENTIALLY AMYLOIDOGENIC, CARBOXYL-TERMINAL DERIVATIVES OF THE AMYLOID PROTEIN-PRECURSOR [J].
ESTUS, S ;
GOLDE, TE ;
KUNISHITA, T ;
BLADES, D ;
LOWERY, D ;
EISEN, M ;
USIAK, M ;
QU, XM ;
TABIRA, T ;
GREENBERG, BD ;
YOUNKIN, SG .
SCIENCE, 1992, 255 (5045) :726-728