title: Proximity-Dependent Biotinylation for Identification of Interacting Proteins.
abstract: Complex interaction networks orchestrate key cellular processes including but not limited to transcription, translation, metabolism, and cell signaling. Delineating these interactions will aid in deciphering the regulation and function of these pathways and potential for manipulation. Proximity-dependent biotin identification (BioID) is quickly gaining popularity as a powerful tool for identifying novel protein-protein and proximity-based interactions in live cells. This technique relies on a promiscuous biotin ligase, which is fused to a protein of interest and, upon expression in the desired cell, will biotinylate proximal endogenous proteins. In vivo protein-protein interactions can be very transient and occur momentarily to facilitate signaling or a metabolic function. BioID is useful in identifying these weak and/or transient interactions that are not detected by traditional methods such as yeast two-hybrid or affinity purification. Here, we outline a BioID protocol that can be used as a workflow to guide a new application. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, Québec, Canada.
authors.forename: Valerie
authors.initials: V
authors.lastname: Le Sage
authors.affiliation: HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, Québec, Canada.|Department of Medicine, McGill University, Montréal, Québec, Canada.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Cinti
authors.affiliation: HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital, Montréal, Québec, Canada.|Department of Medicine, McGill University, Montréal, Québec, Canada.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Mouland
mesh_terms: D000818:Animals; D019921:Biotinylation; D002460:Cell Line; D006801:Humans; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D011993:Recombinant Fusion Proteins
keywords: BioID; BirA; protein-protein interactions; proximity biotinylation