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Identification of ATM Protein Kinase Phosphorylation Sites by Mass Spectrometry

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ATM Kinase

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1599))

Abstract

ATM (ataxia-telangiectasia mutated) protein kinase is a key regulator of cellular responses to DNA damage and oxidative stress. DNA damage triggers complex cascade of signaling events leading to numerous posttranslational modification on multitude of proteins. Understanding the regulation of ATM kinase is therefore critical not only for understanding the human genetic disorder ataxia-telangiectasia and potential treatment strategies, but essential for deciphering physiological responses of cells to stress. These responses play an important role in carcinogenesis, neurodegeneration, and aging. We focus here on the identification of DNA damage inducible ATM phosphorylation sites to understand the importance of autophosphorylation in the mechanism of ATM kinase activation. We demonstrate the utility of using immunoprecipitated ATM in quantitative LC-MS/MS workflow with stable isotope dimethyl labeling of ATM peptides for identification of phosphorylation sites.

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Acknowledgments

M.E.G. and M.F.L. were funded by the National Health and Medical Research Council. M.E.G. was funded by the Brain Foundation and equipment grants from the Cancer Institute New South Wales, the Australian Cancer Research Foundation, and Rebecca L Cooper Medical Research Foundation.

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Authors and Affiliations

  1. Children’s Medical Research Institute, University of Sydney, Westmead, NSW, 2145, Australia

    Mark E. Graham

  2. University of Queensland Centre for Clinical Research (UQCCR), University of Queensland, Building 71/918, Royal Brisbane & Women’s Hospital Campus, Herston, Brisbane, QLD4029, Australia

    Martin F. Lavin & Sergei V. Kozlov

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  1. Mark E. Graham
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  2. Martin F. Lavin
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  3. Sergei V. Kozlov
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Correspondence to Sergei V. Kozlov .

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Editors and Affiliations

  1. University of Queensland Centre for Clinical Research (UQCCR), University of Queensland, Herston, Queensland, Australia

    Sergei V. Kozlov

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Graham, M.E., Lavin, M.F., Kozlov, S.V. (2017). Identification of ATM Protein Kinase Phosphorylation Sites by Mass Spectrometry. In: Kozlov, S. (eds) ATM Kinase. Methods in Molecular Biology, vol 1599. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6955-5_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6955-5_10

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  • Print ISBN: 978-1-4939-6953-1

  • Online ISBN: 978-1-4939-6955-5

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