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Development of Selected Reaction Monitoring Methods to Systematically Quantify Kinase Abundance and Phosphorylation Stoichiometry in Human Samples

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Kinase Signaling Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1636))

Abstract

Protein phosphorylation, one of the most common types of post-translational modifications, is the central regulatory mechanism of cellular signaling networks. In human cells, thousands of proteins are continuously and dynamically phosphorylated and dephosphorylated at specific sites and times in response to external and internal stimuli. Reversible phosphorylation is facilitated by the action of two protein superfamilies: kinases and phosphatases. Kinases play an essential role in almost every relevant physiological process in human cells and their deregulation is linked to pathologies ranging from cancer to autoimmune diseases.

Systematic identification of kinases expressed in a particular cell type, quantification of their abundance, and precise determination of their phosphorylation stoichiometry are essential to understand the cellular signaling networks and physiology of a sample. Our protocol outlines the steps to build and use a high-throughput, comprehensive, modular, and robust selected reaction monitoring (SRM) proteomics framework to facilitate quantification of the kinome state in research or clinical human samples.

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Author information

Author notes

  1. Kirsten Beck and Nathan Camp contributed equally to this work.

Authors and Affiliations

  1. Department of Genome Sciences, University of Washington, Seattle, WA, 98115, USA

    Kirsten Beck, Michael Bereman, James Bollinger, Jarrett Egertson, Michael MacCoss & Alejandro Wolf-Yadlin

  2. Quintara Biosciences, San Francisco, CA, 94080, USA

    Kirsten Beck

  3. North Carolina State University, Raleigh, NC, 27695, USA

    Michael Bereman

  4. Washington University, St. Louis, MO, 63130, USA

    James Bollinger

  5. Center for Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA

    Nathan Camp

Authors
  1. Kirsten Beck
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  2. Nathan Camp
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  3. Michael Bereman
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  4. James Bollinger
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  5. Jarrett Egertson
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  6. Michael MacCoss
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  7. Alejandro Wolf-Yadlin
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Corresponding author

Correspondence to Alejandro Wolf-Yadlin .

Editor information

Editors and Affiliations

  1. Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA

    Aik-Choon Tan

  2. Protein Networks Laboratory, Institute of Cancer Research, London, United Kingdom

    Paul H. Huang

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Beck, K. et al. (2017). Development of Selected Reaction Monitoring Methods to Systematically Quantify Kinase Abundance and Phosphorylation Stoichiometry in Human Samples. In: Tan, AC., Huang, P. (eds) Kinase Signaling Networks. Methods in Molecular Biology, vol 1636. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7154-1_23

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  • DOI: https://doi.org/10.1007/978-1-4939-7154-1_23

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7152-7

  • Online ISBN: 978-1-4939-7154-1

  • eBook Packages: Springer Protocols

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