Affinity‐purification coupled to mass spectrometry: Basic principles and strategies

Identifying the interactions established by a protein of interest can be a critical step in understanding its function. This is especially true when an unknown protein of interest is demonstrated to physically interact with proteins of known function. While many techniques have been developed to characterize protein–protein interactions, one strategy that has gained considerable momentum over the past decade for identification and quantification of protein–protein interactions, is affinity‐purification followed by mass spectrometry ( AP ‐ MS ). Here, we briefly review the basic principles used in affinity‐purification coupled to mass spectrometry, with an emphasis on tools (both biochemical and computational), which enable the discovery and reporting of high quality protein–protein interactions.