Characterization of the ectodomain shedding of the beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1)

J Biol Chem. 2003 Sep 19;278(38):36264-8. doi: 10.1074/jbc.M304186200. Epub 2003 Jul 11.

Abstract

Generation of the amyloid peptide through proteolytic processing of the amyloid precursor protein by beta- and gamma-secretases is central to the etiology of Alzheimer's disease. beta-secretase, known more widely as the beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), has been identified as a transmembrane aspartic proteinase, and its ectodomain has been reported to be cleaved and secreted from cells in a soluble form. The extracellular domains of many diverse proteins are known to be cleaved and secreted from cells by a process known as ectodomain shedding. Here we confirm that the ectodomain of BACE1 is secreted from cells and that this processing is up-regulated by agents that activate protein kinase C. A metalloproteinase is involved in the cleavage of BACE1 as hydroxamic acid-based metalloproteinase inhibitors abolish the release of shed BACE1. Using potent and selective inhibitors, we demonstrate that ADAM10 is a strong candidate for the BACE1 sheddase. In addition, we show that the BACE1 sheddase is distinct from alpha-secretase and, importantly, that inhibition of BACE1 shedding does not influence amyloid precursor protein processing at the beta-site.

MeSH terms

  • Amyloid Precursor Protein Secretases
  • Aspartic Acid Endopeptidases / chemistry*
  • Binding Sites
  • Blotting, Western
  • Cell Line
  • Dose-Response Relationship, Drug
  • Electrophoresis, Polyacrylamide Gel
  • Endopeptidases / chemistry
  • Enzyme Activation
  • Humans
  • Hydroxamic Acids / chemistry
  • Inhibitory Concentration 50
  • Models, Chemical
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Kinase C / metabolism
  • Protein Structure, Tertiary
  • Up-Regulation

Substances

  • Hydroxamic Acids
  • Protein Kinase C
  • Amyloid Precursor Protein Secretases
  • Endopeptidases
  • Aspartic Acid Endopeptidases
  • BACE1 protein, human