Unit

UNIT 19.23 BioID: A Screen for Protein-Protein Interactions

  1. Kyle J. Roux1,2,
  2. Dae In Kim1,
  3. Brian Burke3

Published Online: 5 NOV 2013

DOI: 10.1002/0471140864.ps1923s74

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Roux, K. J., Kim, D. I. and Burke, B. 2013. BioID: A Screen for Protein-Protein Interactions. Current Protocols in Protein Science. 74:19.23:19.23.1–19.23.14.

Author Information

  1. 1

    Children's Health Research Center, Sanford Research, North Sioux Falls, South Dakota

  2. 2

    Department of Pediatrics, Sanford School of Medicine, University of South Dakota, Sioux Falls, South Dakota

  3. 3

    Institute of Medical Biology, Immunos, Singapore

Publication History

  1. Published Online: 5 NOV 2013

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. Curr. Protoc. Protein Sci. 74:19.23.1-19.23.14. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • BioID;
  • proximity-dependent labeling;
  • protein-protein interaction;
  • biotinylation