Format

Send to

Choose Destination
Methods Mol Biol. 2016;1450:35-44. doi: 10.1007/978-1-4939-3759-2_4.

Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.

Author information

1
Department of Plant Systems Biology, Technische Universität München, Emil-Ramann-Str. 4, 85354, Freising, Germany.
2
Department of Plant Systems Biology, Technische Universität München, Emil-Ramann-Str. 4, 85354, Freising, Germany. erika.isono@wzw.tum.de.

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.

KEYWORDS:

DUB assay; Deubiquitinating enzymes; K48-linked ubiquitin chains; K63-linked ubiquitin chains

PMID:
27424744
DOI:
10.1007/978-1-4939-3759-2_4
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center