|Publication Type||Journal Article|
|Year of Publication||2009|
|Authors||Ong, SE, Schenone, M, Margolin, AA, Li, X, Do, K, Doud, MK, Mani, DR, Kuai, L, Wang, X, Wood, JL, Tolliday, NJ, Koehler, AN, Marcaurelle, LA, Golub, TR, Gould, RJ, Schreiber, SL, Carr, SA|
|Journal||Proceedings of the National Academy of Sciences of the United States of America|
Most small-molecule probes and drugs alter cell circuitry by interacting with 1 or more proteins. A complete understanding of the interacting proteins and their associated protein complexes, whether the compounds are discovered by cell-based phenotypic or target-based screens, is extremely rare. Such a capability is expected to be highly illuminating--providing strong clues to the mechanisms used by small-molecules to achieve their recognized actions and suggesting potential unrecognized actions. We describe a powerful method combining quantitative proteomics (SILAC) with affinity enrichment to provide unbiased, robust and comprehensive identification of the proteins that bind to small-molecule probes and drugs. The method is scalable and general, requiring little optimization across different compound classes, and has already had a transformative effect on our studies of small-molecule probes. Here, we describe in full detail the application of the method to identify targets of kinase inhibitors and immunophilin binders.