Identifying the proteins to which small-molecule probes and drugs bind in cells
Author(s)
Ong, Shao-En; Schenone, Monica; Margolin, Adam A.; Li, Xiaoyu; Do, Kathy; Doud, Mary Kathryn; Mani, D. R.; Kuai, Letian; Wang, Xiang; Wood, John L.; Tolliday, Nicola J.; Koehler, Angela Nicole; Marcaurelle, Lisa A.; Golub, Todd R.; Gould, Robert J.; Schreiber, Stuart L.; Carr, Steven A.; ... Show more Show less
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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.
Date issued
2009-03Department
Broad Institute of MIT and HarvardJournal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
National Academy of Sciences
Citation
Ong, Shao-En et al. “Identifying the proteins to which small-molecule probes and drugs bind in cells.” Proceedings of the National Academy of Sciences 106.12 (2009): 4617-4622.
Version: Final published version
ISSN
0027-8424