Ong SE, Schenone M, Margolin AA, Li X, Do K, Doud MK, Mani DR, Kuai L, Wang X, Wood JL, et al. Identifying the proteins to which small-molecule probes and drugs bind in cells. Proc Natl Acad Sci U S A. 2009;106:4617–22.
NOTES
Ong, Shao-EnSchenone, MonicaMargolin, Adam ALi, XiaoyuDo, KathyDoud, Mary KMani, D RKuai, LetianWang, XiangWood, John LTolliday, Nicola JKoehler, Angela NMarcaurelle, Lisa AGolub, Todd RGould, Robert JSchreiber, Stuart LCarr, Steven AengUL1RR024924/RR/NCRR NIH HHS/RL1HG004671/HG/NHGRI NIH HHS/RL1CA133834/CA/NCI NIH HHS/RL1GM084437/GM/NIGMS NIH HHS/UL1 RR024924/RR/NCRR NIH HHS/RL1 GM084437/GM/NIGMS NIH HHS/N01CO12400/CA/NCI NIH HHS/RL1 HG004671/HG/NHGRI NIH HHS/RL1 CA133834/CA/NCI NIH HHS/N01-CO-12400/CO/NCI NIH HHS/Research Support, N.I.H., ExtramuralProc Natl Acad Sci U S A. 2009 Mar 24;106(12):4617-22. doi: 10.1073/pnas.0900191106. Epub 2009 Mar 2.
Abstract
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.
Last updated on 02/17/2021