Context-Specific and Proximity-Dependent Labeling for the Proteomic Analysis of Spatiotemporally Defined Protein Complexes with Split-BioID

Methods Mol Biol. 2021:2247:303-318. doi: 10.1007/978-1-0716-1126-5_17.

Abstract

Proximity-dependent labeling techniques such as BioID and APEX2 allow the biotinylation of proteins proximal to a protein of interest in living cells. Following streptavidin pulldown and mass spectrometry analysis, this enables the identification of native protein-protein interactions. Here we describe split-BioID, a protein-fragment complementation assay that increases the resolution of BioID. Using this technique, context-specific protein complexes can be resolved.

Keywords: BioID; Biotin; Protein-fragment complementation assay; Protein–protein interactions; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotinylation
  • Cell Line
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression
  • Humans
  • Mass Spectrometry
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Proteins / genetics
  • Proteins / metabolism
  • Proteome*
  • Proteomics / methods*
  • Staining and Labeling
  • Streptavidin / metabolism

Substances

  • Proteins
  • Proteome
  • Streptavidin