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Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes

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Book cover Plant Proteostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1450))

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Abstract

Deubiquitylating enzymes, or DUBs, are important regulators of ubiquitin homeostasis and substrate stability, though the molecular mechanisms of most of the DUBs in plants are not yet understood. As different ubiquitin chain types are implicated in different biological pathways, it is important to analyze the enzyme characteristic for studying a DUB. Quantitative analysis of DUB activity is also important to determine enzyme kinetics and the influence of DUB binding proteins on the enzyme activity. Here, we show methods to analyze DUB activity using immunodetection, Coomassie Brilliant Blue staining, and fluorescence measurement that can be useful for understanding the basic characteristic of DUBs.

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Correspondence to Erika Isono .

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Kalinowska, K., Nagel, MK., Isono, E. (2016). Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes. In: Lois, L., Matthiesen, R. (eds) Plant Proteostasis. Methods in Molecular Biology, vol 1450. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3759-2_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3759-2_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3757-8

  • Online ISBN: 978-1-4939-3759-2

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