Identifying the proteins to which small-molecule probes and drugs bind in cells

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