Defining the interactomes of proteins involved in cytoskeletal dynamics using high-throughput proximity-dependent biotinylation in cellulo

STAR Protoc. 2022 Jan 10;3(1):101075. doi: 10.1016/j.xpro.2021.101075. eCollection 2022 Mar 18.


Proximity-dependent biotinylation (BioID) screens are excellent tools to capture in cellulo interactomes for a large variety of baits, including transient and weak affinity interactions, as well as localization-specific proximity components, which are much harder to detect with conventional approaches. Here, we describe the major starting steps and a detailed protocol on how to perform BioID in mammalian cells. We also describe the mass spectrometry procedure and the bioinformatics pipeline for the data analysis. For complete details on the use and execution of this profile, please refer to Bagci et al. (2020).

Keywords: Bioinformatics; Cell Biology; High Throughput Screening; Mass Spectrometry; Molecular Biology; Protein Biochemistry; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotinylation
  • Computational Biology
  • Mammals / metabolism
  • Mass Spectrometry / methods
  • Protein Interaction Mapping* / methods
  • Proteins* / metabolism


  • Proteins