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Screening for Inhibitors of Kinase Autophosphorylation

  • Bianca Heedmann
  • Martin KlumppEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1439)

Abstract

Autophosphorylation of kinases influences their conformational state and can also regulate enzymatic activity. Recently, this has become an area of interest for drug discovery. Using Alk2 as an example, we present two protocols — one based on phosphate-binding Alphascreen beads, the other on coupled luminescence measurements of ADP formation — that can be used to screen for inhibitors of autophosphorylation.

Key words

Kinase Autoactivation Autophosphorylation Alphascreen Luminescence ADP-Glo Alk2 Assay development High-throughput screening 

Notes

Acknowledgments

We highly appreciate the supply of recombinant Alk2 and its substrate Smad1-MH2 by Sonia Faut and Myriam Duckely as well as protein characterization through LC-MS by Peggy Lefeuvre and Francis Bitsch. The support from Geoffrey Cutler in implementing the miniaturized ADP formation assay on an automated screening system was also very helpful. The whole Alk2 project team is acknowledged for input and stimulating discussions. Last but not least, we would like to thank Joerg Trappe for suggesting and encouraging the drafting of this manuscript.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Center for Proteomic ChemistryNovartis Institute for Biomedical Research BaselBaselSwitzerland

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