title: Purification and cloning of amyloid precursor protein beta-secretase from human brain.
abstract: Proteolytic processing of the amyloid precursor protein (APP) generates amyloid beta (Abeta) peptide, which is thought to be causal for the pathology and subsequent cognitive decline in Alzheimer's disease. Cleavage by beta-secretase at the amino terminus of the Abeta peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated carboxy-terminal fragment. Cleavage of the C-terminal fragment by gamma-secretase(s) leads to the formation of Abeta. The pathogenic mutation K670M671-->N670L671 at the beta-secretase cleavage site in APP, which was discovered in a Swedish family with familial Alzheimer's disease, leads to increased beta-secretase cleavage of the mutant substrate. Here we describe a membrane-bound enzyme activity that cleaves full-length APP at the beta-secretase cleavage site, and find it to be the predominant beta-cleavage activity in human brain. We have purified this enzyme activity to homogeneity from human brain using a new substrate analogue inhibitor of the enzyme activity, and show that the purified enzyme has all the properties predicted for beta-secretase. Cloning and expression of the enzyme reveals that human brain beta-secretase is a new membrane-bound aspartic proteinase.
authors.affiliation: Elan Pharmaceuticals, South San Francisco, California 94080, USA. ssinha@elanpharama.com
authors.forename: S
authors.initials: S
authors.lastname: Sinha
authors.forename: J P
authors.initials: JP
authors.lastname: Anderson
authors.forename: R
authors.initials: R
authors.lastname: Barbour
authors.forename: G S
authors.initials: GS
authors.lastname: Basi
authors.forename: R
authors.initials: R
authors.lastname: Caccavello
authors.forename: D
authors.initials: D
authors.lastname: Davis
authors.forename: M
authors.initials: M
authors.lastname: Doan
authors.forename: H F
authors.initials: HF
authors.lastname: Dovey
authors.forename: N
authors.initials: N
authors.lastname: Frigon
authors.forename: J
authors.initials: J
authors.lastname: Hong
authors.forename: K
authors.initials: K
authors.lastname: Jacobson-Croak
authors.forename: N
authors.initials: N
authors.lastname: Jewett
authors.forename: P
authors.initials: P
authors.lastname: Keim
authors.forename: J
authors.initials: J
authors.lastname: Knops
authors.forename: I
authors.initials: I
authors.lastname: Lieberburg
authors.forename: M
authors.initials: M
authors.lastname: Power
authors.forename: H
authors.initials: H
authors.lastname: Tan
authors.forename: G
authors.initials: G
authors.lastname: Tatsuno
authors.forename: J
authors.initials: J
authors.lastname: Tung
authors.forename: D
authors.initials: D
authors.lastname: Schenk
authors.forename: P
authors.initials: P
authors.lastname: Seubert
authors.forename: S M
authors.initials: SM
authors.lastname: Suomensaari
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: D
authors.initials: D
authors.lastname: Walker
authors.forename: J
authors.initials: J
authors.lastname: Zhao
authors.forename: L
authors.initials: L
authors.lastname: McConlogue
authors.forename: V
authors.initials: V
authors.lastname: John
mesh_terms: D000595:Amino Acid Sequence; D053829:Amyloid Precursor Protein Secretases; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D006224:Cricetinae; D010450:Endopeptidases; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014162:Transfection
chemical_list: D016564:Amyloid beta-Protein Precursor; D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509284:BACE2 protein, human; C509280:BACE1 protein, human; C509282:Bace1 protein, mouse