Quantitative interaction proteomics reveals differences in the interactomes of amyloid precursor protein isoforms

Robert J Andrew, Kate Fisher, Kate J Heesom, Katherine A B Kellett, Nigel M Hooper

Research output: Contribution to journalArticle (Academic Journal)peer-review

2 Citations (Scopus)
103 Downloads (Pure)

Abstract

The generation of the amyloid‐β (Aβ) peptides from the amyloid precursor protein (APP) through sequential proteolysis by β‐ and γ‐secretases is a key pathological event in the initiation and propagation of Alzheimer's disease. Aβ and the transcriptionally active APP intracellular domain are generated preferentially from the APP695 isoform compared to the longer APP751 isoform. As the Aβ and amyloid precursor protein intracellular domain produced from cleavage of APP695 and APP751 are identical we hypothesised that the two isoforms have differences within their interactomes which mediate the differential processing of the two isoforms. To investigate this, we applied a proteomics‐based approach to identify differences in the interactomes of the APP695 and APP751 isoforms. Using stable isotope labelling of amino acids in cell culture and quantitative proteomics, we compared the interactomes of APP695 and APP751 expressed in human SH‐SY5Y cells. Through this approach, we identified enrichment of proteins involved in mitochondrial function, the nuclear pore and nuclear transport specifically in the APP695 interactome. Further interrogation of the APP interactome and subsequent experimental validation (co‐immunoprecipitation and siRNA knockdown) revealed GAP43 as a specific modulator of APP751 proteolysis, altering Aβ generation. Our data indicate that interrogation of the APP interactome can be exploited to identify proteins which influence APP proteolysis and Aβ production in an isoform dependent‐manner.

Original languageEnglish
Pages (from-to)399-412
Number of pages14
JournalJournal of Neurochemistry
Volume149
Issue number3
Early online date21 Jan 2019
DOIs
Publication statusPublished - 1 May 2019

Keywords

  • amyloid precursor protein
  • Amyloid-β
  • GAP43
  • interactome
  • proteolysis
  • SILAC

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