Purification and cloning of amyloid precursor protein β-secretase from human brain

• Published in: Nature (London) Vol. 402; no. 6761; pp. 537 - 540
• Main Authors: Sinha, Sukanto, Anderson, John P, Barbour, Robin, Basi, Guriqbal S, Caccavello, Russell, Davis, David, Doan, Minhtam, Dovey, Harry F, Frigon, Normand, Hong, Jin, Jacobson-Croak, Kirsten, Jewett, Nancy, Keim, Pamela, Knops, Jeroen, Lieberburg, Ivan, Power, Michael, Tan, Hua, Tatsuno, Gwen, Tung, Jay, Schenk, Dale, Seubert, Peter, Suomensaari, Susanna M, Wang, Shuwen, Walker, Donald, Zhao, Jun, McConlogue, Lisa, John, Varghese
• Format: Journal Article
• Language: English
• Published: London Nature Publishing 02.12.1999; Nature Publishing Group
• Subjects: Alzheimer's disease; Amino Acid Sequence; Amyloid beta-Protein Precursor - metabolism; Amyloid Precursor Protein Secretases; Animals; Aspartic Acid Endopeptidases - antagonists & inhibitors; Aspartic Acid Endopeptidases - genetics; Aspartic Acid Endopeptidases - isolation & purification; Aspartic Acid Endopeptidases - metabolism; aspartic acid proteinase; b-secretase; Biological and medical sciences; Brain; Brain - enzymology; Cell Line; Cell Membrane - enzymology; CHO Cells; Cloning; Cloning, Molecular; Cricetinae; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; Endopeptidases; Enzyme Inhibitors - pharmacology; Enzymes; Escherichia coli; Humans; Medical sciences; Membranes; Mice; Molecular Sequence Data; Mutation; Neurology; Peptides; Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Sequence Homology, Amino Acid; Tissue Distribution; Transfection; Human; Nervous system diseases; Purification; Nucleotide sequence; Enzyme; Alzheimer disease; Homology; Gene expression; Amyloid precursor protein; Cerebral disorder; Peptidases; Enzymatic activity; Gene; Proteolysis; Central nervous system disease; Hydrolases; Genetics; Degenerative disease; Aspartic endopeptidases
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/990114

Proteolytic processing of the amyloid precursor protein (APP) generates amyloid β (Aβ) peptide, which is thought to be causal for the pathology and subsequent cognitive decline in Alzheimer's disease. Cleavage by β-secretase at the amino terminus of the Aβ peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of β-cleaved soluble APP, and a corresponding cell-associated carboxy-terminal fragment. Cleavage of the C-terminal fragment by γ-secretase(s) leads to the formation of Aβ. The pathogenic mutation K670M671 → N670L671 at the β-secretase cleavage site in APP, which was discovered in a Swedish family with familial Alzheimer's disease, leads to increased β-secretase cleavage of the mutant substrate. Here we describe a membrane-bound enzyme activity that cleaves full-length APP at the β-secretase cleavage site, and find it to be the predominant β-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 β-secretase. Cloning and expression of the enzyme reveals that human brain β-secretase is a new membrane-bound aspartic proteinase.