A high-throughput approach for measuring temporal changes in the interactome

Interactomes are often measured using affinity purification-mass spectrometry (AP-MSMS) or yeast two-hybrid approaches, but these methods do not provide stoichiometric or temporal information. We combine quantitative proteomics and size-exclusion chromatography to map 291 coeluting complexes. This method allows mapping of an interactome to the same depth and accuracy as AP-MSMS with less work and without overexpression or tagging. The use of triplex labeling enables monitoring of interactome rearrangements.