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

Interactomes are often measured using affinity purification-mass spectrometry (AP-MS) or yeast two-hybrid approaches but these lack 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-MS with less work and without overexpression or tagging. The use of triplex labeling enables monitoring of interactome rearrangements.