title: A synthetic substrate assay for the gamma-secretase of the beta-A4 amyloid of Alzheimer's disease.
abstract: gamma-secretase, the endoprotease which releases the C-terminus of beta A4 amyloid peptide, cleaves within the hydrophobic transmembrane domain of the amyloid precursor protein. In order to obtain a substrate for gamma-secretase, a dodecapeptide which spans the cleavage site was synthesized, labelled with 125-iodine and conjugated to an agarose gel. A radiometric solid-phase assay was developed using this immobilized substrate. Peptide products were separated by reverse-phase HPLC and TLC to allow characterization of the cleavage site(s).
authors.affiliation: Department of Pathology, University of Melbourne, Australia.
authors.forename: G
authors.initials: G
authors.lastname: Evin
authors.forename: K
authors.initials: K
authors.lastname: Beyreuther
authors.forename: C L
authors.initials: CL
authors.lastname: Masters
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism*; D000595:Amino Acid Sequence; D053829:Amyloid Precursor Protein Secretases; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016282:Aspartic Acid Endopeptidases; D001665:Binding Sites / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D010450:Endopeptidases / Q000032:analysis* / Q000378:metabolism*; D006801:Humans; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity
chemical_list: D016229:Amyloid beta-Peptides; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human