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The Hippo Pathway pp 289–304Cite as

Quantifying the Kinase Activities of MST1/2

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1893))

Abstract

The functions of the kinases MST1 and MST2 rely heavily on their ability to phosphorylate and become phosphorylated themselves. Hence, it is important to precisely measure the kinase activities of both isoforms in a reproducible manner. Here, we describe in detail the protocol for an in-gel kinase assay for the quantification of the kinase activity of MST1/2, which involves immunoprecipitation of MST1/2 and the incorporation of radiolabeled phosphate from [γ-32P]-ATP into a substrate immobilized in a polyacrylamide gel. We also include a protocol for indirect measurement of MST1/2 activation status using immunoblotting.

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Acknowledgments

This work was supported by Science Foundation Ireland grants 15/CDA/3495 and 16/TIDA/4064.

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Authors and Affiliations

  1. Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland

    Niamh A. O’Driscoll & David Matallanas

  2. School of Medicine, University College Dublin, Belfield, Dublin, Ireland

    Niamh A. O’Driscoll & David Matallanas

Authors
  1. Niamh A. O’Driscoll
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  2. David Matallanas
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Corresponding author

Correspondence to David Matallanas .

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Editors and Affiliations

  1. Cancer Institute, University College London, London, UK

    Alexander Hergovich

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O’Driscoll, N.A., Matallanas, D. (2019). Quantifying the Kinase Activities of MST1/2. In: Hergovich, A. (eds) The Hippo Pathway. Methods in Molecular Biology, vol 1893. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8910-2_22

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  • DOI: https://doi.org/10.1007/978-1-4939-8910-2_22

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8909-6

  • Online ISBN: 978-1-4939-8910-2

  • eBook Packages: Springer Protocols

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