EXPRESSION OF HUMAN ALZHEIMER AMYLOID PRECURSOR PROTEIN IN INSECT CELLS

被引:23
作者
RAMAKRISHNA, N [1 ]
SAIKUMAR, P [1 ]
POTEMPSKA, A [1 ]
WISNIEWSKI, HM [1 ]
MILLER, DL [1 ]
机构
[1] WISTAR INST,PHILADELPHIA,PA 19104
关键词
D O I
10.1016/0006-291X(91)91515-E
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
The amyloid beta-peptide is a major constituent of amyloid deposited in the brains of patients with Alzheimer's disease and is derived from a larger precursor protein/s (APP-695, 751, 770). A human cDNA encoding full-length APP-751 was inserted into the genome of Autographa californica nuclear polyhedrosis virus under transcriptional regulation of the viral polyhedrin gene promoter. The recombinant virus was used to infect insect cells, which resulted in the abundant expression of APP-751. Analysis of infected cell proteins indicate that APP-751 is localized in the membrane fraction; however, a significant amount of the protein was cleaved and released into the medium. The NH2-terminal sequence of recombinant APP-751 from the membrane fraction was identical to that of mammalian APP. Immunoblot analysis suggests that the secreted form results from cleavage within the β-peptide. © 1991.
引用
收藏
页码:983 / 989
页数:7
相关论文
共 25 条
[1]   TRANSFORMING GROWTH-FACTOR-BETA BOUND TO SOLUBLE DERIVATIVES OF THE BETA-AMYLOID PRECURSOR PROTEIN OF ALZHEIMERS-DISEASE [J].
BODMER, S ;
PODLISNY, MB ;
SELKOE, DJ ;
HEID, I ;
FONTANA, A .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1990, 171 (02) :890-897
[2]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[3]   SITE-SPECIFIC MUTATION OF THE CONSERVED M-2(6)A M-2(6)A RESIDUES OF ESCHERICHIA-COLI 16S RIBOSOMAL-RNA - EFFECTS ON RIBOSOME FUNCTION AND ACTIVITY OF THE KSGA METHYLTRANSFERASE [J].
CUNNINGHAM, PR ;
WEITZMANN, CJ ;
NURSE, K ;
MASUREL, R ;
VANKNIPPENBERG, PH ;
OFENGAND, J .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1050 (1-3) :18-26
[4]  
DENMAN RB, 1990, UNPUB
[5]  
ESCH FS, 1990, SCIENCE, V248, P1112
[6]   A HIGH-YIELD METHOD FOR THE ISOLATION OF HYDROPHOBIC PROTEINS AND PEPTIDES FROM POLYACRYLAMIDE GELS FOR PROTEIN SEQUENCING [J].
FEICK, RG ;
SHIOZAWA, JA .
ANALYTICAL BIOCHEMISTRY, 1990, 187 (02) :205-211
[7]   STRUCTURE OF PREPROATTACIN AND ITS PROCESSING IN INSECT CELLS INFECTED WITH A RECOMBINANT BACULOVIRUS [J].
GUNNE, H ;
HELLERS, M ;
STEINER, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 187 (03) :699-703
[8]   THE PRECURSOR OF ALZHEIMERS-DISEASE AMYLOID-A4 PROTEIN RESEMBLES A CELL-SURFACE RECEPTOR [J].
KANG, J ;
LEMAIRE, HG ;
UNTERBECK, A ;
SALBAUM, JM ;
MASTERS, CL ;
GRZESCHIK, KH ;
MULTHAUP, G ;
BEYREUTHER, K ;
MULLERHILL, B .
NATURE, 1987, 325 (6106) :733-736
[9]  
Kim K. S., 1988, NEUROSCI RES COMMUN, V2, P121
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+