BioID: a screen for protein-protein interactions

Curr Protoc Protein Sci. 2013 Nov 5;74:19.23.1-19.23.14. doi: 10.1002/0471140864.ps1923s74.

Abstract

BioID is a unique method to screen for physiologically relevant protein interactions that occur in living cells. This technique harnesses a promiscuous biotin ligase to biotinylate proteins based on proximity. The ligase is fused to a protein of interest and expressed in cells, where it biotinylates proximal endogenous proteins. Because it is a rare protein modification in nature, biotinylation of these endogenous proteins by BioID fusion proteins enables their selective isolation and identification with standard biotin-affinity capture. Proteins identified by BioID are candidate interactors for the protein of interest. BioID can be applied to insoluble proteins, can identify weak and/or transient interactions, and is amenable to temporal regulation. Initially applied to mammalian cells, BioID has potential application in a variety of cell types from diverse species.

Keywords: BioID; biotinylation; protein-protein interaction; proximity-dependent labeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotinylation / methods*
  • Immunoblotting
  • Protein Interaction Mapping / methods*
  • Proteins / analysis*
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism*
  • Transfection

Substances

  • Proteins