Proteolysis of chimeric β-amyloid precursor proteins containing the Notch transmembrane domain yields amyloid β-like peptides

Jimin Zhang, Wenjuan Ye, Rong Wang, Michael S. Wolfe, Barry D. Greenberg, Dennis J. Selkoe

Research output: Contribution to journalArticlepeer-review

Abstract

γ-Secretase is an unusual intramembranous protease that has been reported to cleave the β-amyloid precursor protein (APP) near the middle of its transmembrane domain (TMD) but cleave Notch near the cytoplasmic end of its TMD. To ascertain whether the TMD sequence of the substrate determines where γ-secretase cleaves and whether the region just before the TMD participates in recognition by the enzyme, we expressed chimeric human APP molecules containing either the TMD or pre-TMD regions of Notch or other transmembrane proteins. APP chimeras bearing either the Notch or the amyloid precursor-like protein-2 TMD released similar amounts of ∼4-kDa amyloid β-peptide (Aβ)-like peptides as did intact APP. Mass spectrometry revealed that the principal Aβ-like peptide ended at residue 40, indicating cleavage at the middle of the Notch TMD in the chimera. Generation of Aβ-like peptides was significantly decreased when the APP TMD was replaced by those of SREBP-1 or human epithelial growth factor receptor 3. Replacement of the APP pre-TMD region (Aβ 10-28) with that of SREBP-1 increased generation of Aβ-like peptides, while those of human epithelial growth factor receptor 3 or amyloid precursor-like protein-2 decreased it. We conclude that γ-secretase can cleave near the middle of the Notch TMD, that Aβ-like peptides may arise during Notch processing, and that the pre-TMD sequence of the substrate influences recognition or binding by the enzyme.

Original languageEnglish (US)
Pages (from-to)15069-15075
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number17
DOIs
StatePublished - Apr 26 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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