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PMID: 20534834 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Three-amino acid spacing of presenilin endoproteolysis suggests a general stepwise cleavage of gamma-secretase-mediated intramembrane proteolysis.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 30 ·No. 23 ·2010-06-09 ·Pages 7853-62

Fukumori A, Fluhrer R, Steiner H, Haass C

Abstract

Presenilin (PS1 or PS2) is the catalytic component of the gamma-secretase complex, which mediates the final proteolytic processing step leading to the Alzheimer's disease (AD)-characterizing amyloid beta-peptide. PS is cleaved during complex assembly into its characteristic N- and C-terminal fragments. Both fragments are integral components of physiologically active gamma-secretase and harbor the two critical aspartyl residues of the active site domain. While the minimal subunit composition of gamma-secretase has been defined and numerous substrates were identified, the cellular mechanism of the endoproteolytic cleavage of PS is still unclear. We addressed this pivotal question by investigating whether familial AD (FAD)-associated PS1 mutations affect the precision of PS endoproteolysis in a manner similar to the way that such mutations shift the intramembrane cleavage of gamma-secretase substrates. We demonstrate that all FAD mutations investigated still allow endoproteolysis to occur. However, the precision of PS1 endoproteolysis is affected by PS1 mutations. Comparing the cleavage products generated by a variety of PS1 mutants revealed that specifically cleavages at positions 293 and 296 of PS1 are selectively affected. Systematic mutagenesis around the cleavage sites revealed a stepwise three amino acid spaced cleavage mechanism of PS endoproteolysis reminiscent to the epsilon-, zeta-, and gamma-cleavages described for typical gamma-secretase substrates, such as the beta-amyloid precursor protein. Our findings therefore suggest that intramembranous cleavage by gamma-secretase and related intramembrane-cleaving proteases may generally occur via stepwise endoproteolysis.

MeSH Terms
Alzheimer Disease/enzymology Amyloid Precursor Protein Secretases/metabolism Amyloid beta-Protein Precursor/metabolism Animals Cell Line Cells, Cultured Humans Immunoblotting Mass Spectrometry Mice Mice, Knockout Mutation Peptide Fragments/metabolism Presenilin-1/genetics,metabolism Presenilin-2/genetics,metabolism Transfection
Chemicals
Amyloid beta-Protein Precursor Peptide Fragments Presenilin-1 Presenilin-2 Amyloid Precursor Protein Secretases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Fukumori Akio
German Center for Neurodegenerative Diseases, Munich, Germany.
Fluhrer Regina
Steiner Harald
Haass Christian
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Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
DOI
10.1523/JNEUROSCI.1443-10.2010
Published
2010-06-09
Pages
7853-62
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6632680
Subset
IM
External Links
PubMed source DOI: 10.1523/JNEUROSCI.1443-10.2010 PMC Full Text View journal details
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