A synthetic substrate assay for the gamma-secretase of the beta-A4 amyloid of Alzheimer's disease

J Pept Sci. 1995 Mar-Apr;1(2):132-9. doi: 10.1002/psc.310010205.

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / metabolism*
  • Amino Acid Sequence
  • Amyloid Precursor Protein Secretases
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / genetics
  • Amyloid beta-Peptides / metabolism*
  • Aspartic Acid Endopeptidases
  • Binding Sites / genetics
  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • Endopeptidases / analysis*
  • Endopeptidases / metabolism*
  • Humans
  • Iodine Radioisotopes
  • Molecular Sequence Data
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification
  • Substrate Specificity

Substances

  • Amyloid beta-Peptides
  • Iodine Radioisotopes
  • Peptide Fragments
  • Amyloid Precursor Protein Secretases
  • Endopeptidases
  • Aspartic Acid Endopeptidases
  • BACE1 protein, human