Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally defined protein complexes

Nat Commun. 2017 Jun 6;8:15690. doi: 10.1038/ncomms15690.

Abstract

Understanding the function of the thousands of cellular proteins is a central question in molecular cell biology. As proteins are typically part of multiple dynamic and often overlapping macromolecular complexes exerting distinct functions, the identification of protein-protein interactions (PPI) and their assignment to specific complexes is a crucial but challenging task. We present a protein fragments complementation assay integrated with the proximity-dependent biotinylation technique BioID. Activated on the interaction of two proteins, split-BioID is a conditional proteomics approach that allows in a single and simple assay to both experimentally validate binary PPI and to unbiasedly identify additional interacting factors. Applying our method to the miRNA-mediated silencing pathway, we can probe the proteomes of two distinct functional complexes containing the Ago2 protein and uncover the protein GIGYF2 as a regulator of miRNA-mediated translation repression. Hence, we provide a novel tool to study dynamic spatiotemporally defined protein complexes in their native cellular environment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay / methods
  • Biotinylation*
  • Carrier Proteins / metabolism
  • Chromatography, Liquid
  • HeLa Cells
  • Humans
  • Mass Spectrometry
  • Phosphorylation
  • Plasmids / metabolism
  • Principal Component Analysis
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Proteome*
  • Proteomics / methods*
  • Recombinant Proteins / metabolism

Substances

  • Carrier Proteins
  • GIGYF2 protein, human
  • Proteome
  • Recombinant Proteins