Expression of human Alzheimer amyloid precursor protein in insect cells.
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 beta-peptide.
Ramakrishna
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N
Department of Molecular Biology, NYS Institute for Basic Research in Developmental Disabilities, Staten Island 10314.
Saikumar
P
P
Potempska
A
A
Wisniewski
H M
HM
Miller
D L
DL
D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D016564:Amyloid beta-Protein Precursor; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D007313:Insecta; D007700:Kinetics; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014162:Transfection
D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D011498:Protein Precursors; D011994:Recombinant Proteins; D004247:DNA