Research abstract
Article abstract
Nature Biotechnology 25, 1035 - 1044 (2007)
Published online: 26 August 2007 | doi:10.1038/nbt1328
Quantitative chemical proteomics reveals mechanisms of action of clinical ABL kinase inhibitors
Marcus Bantscheff1,3, Dirk Eberhard1,3, Yann Abraham1, Sonja Bastuck1, Markus Boesche1, Scott Hobson1, Toby Mathieson1, Jessica Perrin1, Manfred Raida1, Christina Rau1, Valérie Reader2, Gavain Sweetman1, Andreas Bauer1, Tewis Bouwmeester1, Carsten Hopf1, Ulrich Kruse1, Gitte Neubauer1, Nigel Ramsden2, Jens Rick1, Bernhard Kuster1 & Gerard Drewes1
Abstract
We describe a chemical proteomics approach to profile the interaction of small molecules with hundreds of endogenously expressed protein kinases and purine-binding proteins. This subproteome is captured by immobilized nonselective kinase inhibitors (kinobeads), and the bound proteins are quantified in parallel by mass spectrometry using isobaric tags for relative and absolute quantification (iTRAQ). By measuring the competition with the affinity matrix, we assess the binding of drugs to their targets in cell lysates and in cells. By mapping drug-induced changes in the phosphorylation state of the captured proteome, we also analyze signaling pathways downstream of target kinases. Quantitative profiling of the drugs imatinib (Gleevec), dasatinib (Sprycel) and bosutinib in K562 cells confirms known targets including ABL and SRC family kinases and identifies the receptor tyrosine kinase DDR1 and the oxidoreductase NQO2 as novel targets of imatinib. The data suggest that our approach is a valuable tool for drug discovery.
- Cellzome AG, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
- Cellzome UK Ltd., Chesterford Research Park, Cambridge CB10 1XL, UK.
- These authors contributed equally to this work.
Correspondence to: Gerard Drewes1 e-mail: gerard.drewes@cellzome.com
Correspondence to: Bernhard Kuster1 e-mail: bernhard.kuster@cellzome.com
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