Identifying Protein-Protein Interactions by Proximity Biotinylation with AirID and splitAirID.

ABSTRACT: Proteins frequently function in high-order complexes. Defining protein-protein interactions is essential to acquiring a full understanding of their activity and regulation. Proximity biotinylation has emerged as a highly specific approach to capture transient and stable interactions in living cells or organisms. Proximity biotinylation exploits promiscuous biotinylating enzymes fused to a bait protein, resulting in the biotinylation of adjacent endogenous proteins. Biotinylated interactors are purified under very strict conditions and identified by mass spectrometry to obtain a high-confidence list of candidate binding partners. AirID is a recently described biotin ligase specifically engineered for proximity labeling. This protocol details proximity biotinylation by AirID, using protein c

SUBMITTER: Schaack GA 

PROVIDER: S-EPMC10031415 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature