Identifying Cellular Targets of Small-Molecule Probes and Drugs with Biochemical Enrichment and SILAC View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2012

AUTHORS

Shao-En Ong , Xiaoyu Li , Monica Schenone , Stuart L. Schreiber , Steven A. Carr

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. More... »

PAGES

129-40

References to SciGraph publications

  • 2009-09. Target profiling of small molecules by chemical proteomics in NATURE CHEMICAL BIOLOGY
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  • 2009-03. A small molecule that binds Hedgehog and blocks its signaling in human cells in NATURE CHEMICAL BIOLOGY
  • 2006-12. A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC) in NATURE PROTOCOLS
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  • 2007-11. Target deconvolution strategies in drug discovery in NATURE REVIEWS DRUG DISCOVERY
  • 2004-09. Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics in NATURE BIOTECHNOLOGY
  • 2008-07. Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds in NATURE BIOTECHNOLOGY
  • 2005-10. Mass spectrometry–based proteomics turns quantitative in NATURE CHEMICAL BIOLOGY
  • 2009-05. A practical guide to the MaxQuant computational platform for SILAC-based quantitative proteomics in NATURE PROTOCOLS
  • 2010-05. Thousands of chemical starting points for antimalarial lead identification in NATURE
  • 2007-08. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips in NATURE PROTOCOLS
  • 2004-12. Exploring biology with small organic molecules in NATURE
  • 2006-12. In-gel digestion for mass spectrometric characterization of proteins and proteomes in NATURE PROTOCOLS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/978-1-61779-364-6_9

    DOI

    http://dx.doi.org/10.1007/978-1-61779-364-6_9

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1026376650

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/22065222


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