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A Practical Recipe to Survey Phosphoproteomes

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Shotgun Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1156))

Abstract

The field of cellular signaling is fueled by the discovery of novel protein phosphorylation events. Phosphoproteomics focuses on the large-scale identification and characterization of serine, threonine, and tyrosine phosphorylation of proteins. Phosphopeptide enrichment followed by mass spectrometry has emerged as the most powerful technique for unbiased, discovery-driven analysis by offering high sensitivity, resolution, and speed. Methods for mass spectrometry-based phosphoproteomics analysis have improved substantially over the last decade, making the discipline more approachable to the broader scientific community. Herein we describe the status of the field of phosphoproteomics and provide a robust workflow covering the major aspects of large-scale phosphorylation analysis from phosphopeptide enrichment via IMAC to data analysis.

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Acknowledgements

The authors would like to thank Danielle Swaney, Ricard Rodriguez-Mias, and Daniel Hernandez for their helpful advice. This work was supported in part by NIH/NCI grant R00CA140789 and an Ellison Medical Foundation New Scholar Award (to J.V.). W.C.E. is a recipient of an NSF Graduate Research Fellowship.

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Authors and Affiliations

  1. Department of Genome Sciences, University of Washington, Foege S133C, 3720 15th Ave. NE, Seattle, WA, 98105, USA

    William C. Edelman, Kelsey M. Haas, Joanne I. Hsu, Robert T. Lawrence & Judit Villén

Authors
  1. William C. Edelman
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  2. Kelsey M. Haas
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  3. Joanne I. Hsu
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  4. Robert T. Lawrence
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  5. Judit Villén
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Corresponding author

Correspondence to Judit Villén .

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Editors and Affiliations

  1. Ludwig-Maximilians-University, Munich, Germany

    Daniel Martins-de-Souza

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Edelman, W.C., Haas, K.M., Hsu, J.I., Lawrence, R.T., Villén, J. (2014). A Practical Recipe to Survey Phosphoproteomes. In: Martins-de-Souza, D. (eds) Shotgun Proteomics. Methods in Molecular Biology, vol 1156. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0685-7_26

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  • DOI: https://doi.org/10.1007/978-1-4939-0685-7_26

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0684-0

  • Online ISBN: 978-1-4939-0685-7

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