title: Measuring the Enzyme Activity of Arabidopsis Deubiquitylating Enzymes.
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.
authors.affiliation: Department of Plant Systems Biology, Technische Universität München, Emil-Ramann-Str. 4, 85354, Freising, Germany.
authors.forename: Kamila
authors.initials: K
authors.lastname: Kalinowska
authors.affiliation: Department of Plant Systems Biology, Technische Universität München, Emil-Ramann-Str. 4, 85354, Freising, Germany.
authors.forename: Marie-Kristin
authors.initials: MK
authors.lastname: Nagel
authors.affiliation: Department of Plant Systems Biology, Technische Universität München, Emil-Ramann-Str. 4, 85354, Freising, Germany. erika.isono@wzw.tum.de.
authors.forename: Erika
authors.initials: E
authors.lastname: Isono
mesh_terms: D017360:Arabidopsis / Q000201:enzymology*; D000072017:Deubiquitinating Enzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D025801:Ubiquitin / Q000737:chemistry*
chemical_list: D025801:Ubiquitin; D000072017:Deubiquitinating Enzymes
keywords: DUB assay; Deubiquitinating enzymes; K48-linked ubiquitin chains; K63-linked ubiquitin chains