Abstract
Sequencing of the human genome in the last decade has not yet led to a concomitant increase in the numbers of novel drug targets. While the pharmaceutical industry has invested heavily in improving drugs for existing protein targets, it has not tended toward a similar investment in experimental approaches to identify cellular targets of drugs. It is striking that the targets of numerous widely used FDA-approved drugs remain unknown. The development of robust, unbiased methods for target identification would greatly enhance our understanding the mechanisms-of-action of small molecules. Cell-based phenotypic screens followed by unbiased target identification have the potential to identify novel combinations of small molecules and their protein targets, shed light on drug polypharmacology, and enable unbiased screening approaches to drug discovery. Classical biochemical enrichment with immobilized small molecules has been used for over four decades but has been limited by issues concerning specificity and sensitivity. The application of mass spectrometry-based quantitative proteomics in combination with these affinity reagents has proven to be especially useful in addressing these common issues in affinity purification experiments. We describe the use of SILAC in identifying proteins that bind small-molecule probes and drugs in a cellular context.
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Acknowledgments
The authors gratefully acknowledge the helpful discussions and useful insights of our colleagues in the Chemical Biology Program, Chemical Biology Platform, and Proteomics Platform at the Broad Institute. This research has been funded in part with Federal funds from the National Cancer Institute’s Initiative for Chemical Genetics (Contract No. N01-CO-12400) as well as the NIH grant for Genomics Based Drug Discovery – Target ID project (NIH RL1HG004671), administratively linked to RL1CA133834, RL1GM084437, and UL1RR024924.
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Ong, SE., Li, X., Schenone, M., Schreiber, S.L., Carr, S.A. (2012). Identifying Cellular Targets of Small-Molecule Probes and Drugs with Biochemical Enrichment and SILAC. In: Drewes, G., Bantscheff, M. (eds) Chemical Proteomics. Methods in Molecular Biology, vol 803. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-364-6_9
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