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The Ubiquitin Proteasome System pp 385–400Cite as

Interpreting the Language of Polyubiquitin with Linkage-Specific Antibodies and Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1844))

Abstract

Posttranslational modification of cellular proteins by ubiquitin serves a variety of functions. Among the multitude of ubiquitin substrates, ubiquitin itself is the most prevalent. For many years, the direct detection of polyubiquitin chains attached to cellular substrates was not practical, with cell biologists relegated to indirect approaches involving site-directed mutagenesis or in vitro biochemistry. Recent advances in two technologies—polyubiquitin linkage-specific antibodies and mass spectrometry proteomics, have overcome that limitation. Using one or both of these, the direct analysis of polyubiquitin chain linkages on cellular substrate proteins may be performed. This paper describes the complimentary nature of linkage-specific antibodies and mass spectrometry proteomics for the characterization of complex ubiquitin signals using lessons learned in early development of both technologies.

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Acknowledgments

All authors are employees of Genentech Inc. and shareholders of the Roche group.

Author information

Authors and Affiliations

  1. Department of Structural Biology, Genentech, Inc., South San Francisco, CA, USA

    Marissa L. Matsumoto & Erick R. Castellanos

  2. Department of Microchemistry, Proteomics and Lipidomics, Genentech, Inc., South San Francisco, CA, USA

    Yi Jimmy Zeng & Donald S. Kirkpatrick

Authors
  1. Marissa L. Matsumoto
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  2. Erick R. Castellanos
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  3. Yi Jimmy Zeng
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  4. Donald S. Kirkpatrick
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Corresponding authors

Correspondence to Marissa L. Matsumoto or Donald S. Kirkpatrick .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada

    Thibault Mayor

  2. Department of Chemistry and Biochemistry, University of Nevada Las Vegas, Las Vegas, NV, USA

    Gary Kleiger

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Matsumoto, M.L., Castellanos, E.R., Zeng, Y.J., Kirkpatrick, D.S. (2018). Interpreting the Language of Polyubiquitin with Linkage-Specific Antibodies and Mass Spectrometry. In: Mayor, T., Kleiger, G. (eds) The Ubiquitin Proteasome System. Methods in Molecular Biology, vol 1844. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8706-1_24

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  • DOI: https://doi.org/10.1007/978-1-4939-8706-1_24

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8705-4

  • Online ISBN: 978-1-4939-8706-1

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