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Plant Pattern Recognition Receptors pp 109–121Cite as

In Vitro Ubiquitination Activity Assays in Plant Immune Responses

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1578))

Abstract

Ubiquitination is a central posttranslational modification that impinges on the fate of proteins. While attachment of K48-linked chains onto soluble proteins marks them for proteolysis via the 26S proteasome, mono-ubiquitination or K63-linked chains result in the endocytosis and sorting through the endomembrane system of integral membrane proteins, such as pattern recognition receptors. In vitro ubiquitination assays allow the biochemical analysis of all individual components of the ubiquitination machinery and its potential substrates. Here, we describe how to reconstitute the ubiquitination cascade in vitro and detail different variations of the assay, the required controls and how to interpret the obtained results.

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Acknowledgments

The author would like to acknowledge the funding from the Leibniz Association and the state of Saxony-Anhalt.

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Authors and Affiliations

  1. Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120, Halle (Saale), Germany

    Giulia Furlan & Marco Trujillo

Authors
  1. Giulia Furlan
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  2. Marco Trujillo
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Corresponding author

Correspondence to Marco Trujillo .

Editor information

Editors and Affiliations

  1. Department of Plant Pathology and Microbiology Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Libo Shan

  2. Department of Biochemistry and Biophysics Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Ping He

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Furlan, G., Trujillo, M. (2017). In Vitro Ubiquitination Activity Assays in Plant Immune Responses. In: Shan, L., He, P. (eds) Plant Pattern Recognition Receptors. Methods in Molecular Biology, vol 1578. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6859-6_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6859-6_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6858-9

  • Online ISBN: 978-1-4939-6859-6

  • eBook Packages: Springer Protocols

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