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

Nat Methods. 2012 Sep;9(9):907-9. doi: 10.1038/nmeth.2131. Epub 2012 Aug 5.


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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Amino Acids / metabolism
  • Cell Culture Techniques
  • Chromatography, Gel*
  • Chromatography, High Pressure Liquid
  • High-Throughput Screening Assays / methods*
  • Humans
  • Isotope Labeling
  • Protein Interaction Mapping / methods*
  • Protein Interaction Maps*
  • Proteins / analysis
  • Proteins / chemistry
  • Proteins / metabolism*
  • Proteomics / methods*
  • Time Factors


  • Amino Acids
  • Proteins