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Affinity Purification of Proteins Binding to Kinase Inhibitors Immobilized on Self-Assembling Monolayers

  • Marcus Bantscheff
  • Scott Hobson
  • Bernhard KusterEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 795)

Abstract

Kinase inhibitors represent a relatively new class of drugs that offer novel therapies targeting specific ­malfunctioning kinase-mediated signaling pathways in oncology and potentially inflammation. As the ATP binding sites of the ∼500 human kinases are structurally conserved and because most current drugs target the ATP binding site, there is a need to profile all the kinases that a drug may bind and/or inhibit. We have developed a chemical proteomics method that affinity purifies kinases from cell or tissue lysates using kinase inhibitors immobilized on self-assembling monolayers. The method can be applied to assess the selectivity of a given kinase inhibitor and thus to guide its preclinical or clinical development.

Key words

Kinase inhibitor Chemical proteomics Self-assembling monolayers Mass spectrometry 

Notes

Acknowledgments

The authors thank Frank Weisbrodt for help with the figures and Gerard Drewes and Ulrich Kruse for helpful discussions.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Marcus Bantscheff
    • 1
  • Scott Hobson
    • 1
    • 2
  • Bernhard Kuster
    • 3
    Email author
  1. 1.Cellzome AGHeidelbergGermany
  2. 2.Department for CNS ResearchBoehringer Ingelheim GmbH & Co KGBiberach an der RißGermany
  3. 3.TU München LS für ProteomikFreisingGermany

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