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In Vitro Ubiquitination Activity Assays in Plant Immune Responses

Part of the Methods in Molecular Biology book series (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.

Key words

  • Ubiquitination
  • Posttranslational modification
  • Degradation

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  • DOI: 10.1007/978-1-4939-6859-6_8
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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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Correspondence to Marco Trujillo .

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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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