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Chemical proteomics uncovers EPHA2 as a mechanism of acquired resistance to small molecule EGFR kinase inhibition Heiner Koch, M. Estela Del Castillo Busto, Karl Kramer, Guillaume Médard, and Bernhard Kuster J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.5b00161 • Publication Date (Web): 12 May 2015 Downloaded from http://pubs.acs.org on May 13, 2015
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Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
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Journal of Proteome Research
Chemical proteomics uncovers EPHA2 as a mechanism of acquired resistance to small molecule EGFR kinase inhibition Heiner Koch1,2,3,ǁ,M. Estela Del Castillo Busto1,ǁ,#, Karl Kramer1, Guillaume Médard1 and Bernhard Kuster1,2,3,4,5,*
1 Chair for Proteomics and Bioanalytics, Technische Universität München, Freising, Germany 2 German Cancer Consortium (DKTK), Munich, Germany 3 German Cancer Research Center (DKFZ), Heidelberg, Germany 4 Center for Integrated Protein Science Munich, Munich, Germany 5 Bavarian Biomolecular Mass Spectrometry Center, Technische Universität München, Freising, Germany ǁ
These authors contributed equally to this work
#
Current address: Département Biomédical et Chimie Inorganique, Laboratoire national de métrologie
et d'essais (LNE) 1, Paris, France
*Correspondence to: Bernhard Kuster Chair for Proteomics and Bioanalytics, Technische Universität München, Emil Erlenmeyer Forum 5, 85354 Freising, Germany Email:
[email protected] Phone: +49-8161715696 Fax: 49-8161715931
Keywords: kinobeads, chemical proteomics, acquired resistance, kinase inhibitors, EPHA2, EGFR
1 ACS Paragon Plus Environment
Journal of Proteome Research
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Abstract Tyrosine kinase inhibitors (TKIs) have become an important therapeutic option for treating several forms of cancer. Gefitinib, an inhibitor of the epidermal growth factor receptor (EGFR), is in clinical use for treating non-small cell lung cancer (NSCLC) harboring activating EGFR mutations. However, despite high initial response rates, many patients develop resistance to gefitinib. The molecular mechanisms of TKI resistance often remain unclear. Here, we describe a chemical proteomic approach comprising kinase affinity purification (kinobeads) and quantitative mass spectrometry for the identification of kinase inhibitor resistance mechanisms in cancer cells. We identified the previously described amplification of MET and found EPHA2 to be more than 10-fold overexpressed (p
