Large‐scale mapping of human protein–protein interactions by mass spectrometry

Abstract

Mapping protein-protein interactions is an invaluable tool for understanding protein function. Here, we report the first large-scale study of protein-protein interactions in human cells using a mass spectrometry-based approach. The study maps protein interactions for 338 bait proteins that were selected based on known or suspected disease and functional associations. Large-scale immunoprecipitation of Flag-tagged versions of these proteins followed by LC-ESI-MS/MS analysis resulted in the identification of 24 540 potential protein interactions. False positives and redundant hits were filtered out using empirical criteria and a calculated interaction confidence score, producing a data set of 6463 interactions between 2235 distinct proteins. This data set was further cross-validated using previously published and predicted human protein interactions. In-depth mining of the data set shows that it represents a valuable source of novel protein-protein interactions with relevance to human diseases. In addition, via our preliminary analysis, we report many novel protein interactions and pathway associations.

14-3-3 proteins are essential signalling hubs for beta cell survival

“…The differences observed in cell death can be attributed to each isoform having unique interactomes, as structural analysis of the 14-3-3 family revealed a highly variable region between α-helices 8 and 9, which determines binding specificity [42]. This concept has been validated in screens of human 14-3-3 binding partners, which revealed common and unique protein × protein interactions among the tested isoforms [43]. Further work is required to examine the interactome of each 14-3-3 isoform in beta cells.…”

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