title: Development and evaluation of a highly reliable assay for SUMO-specific protease inhibitors.
abstract: SUMOylation, as a post-translational modification of proteins, plays essential regulatory roles in a variety of pathological conditions. In the dynamic process of SUMOylation and deSUMOylation, SENPs (SUMO-specific proteases), in charge of deconjugation of SUMO (small ubiquitin-related modifier) from substrate proteins, have recently been found to be potential therapeutic targets for cancer treatment. A reliable and practical assay is much needed to accelerate the discovery of SENPs inhibitors. We established a quantitative assay based on readily available SDS-PAGE-Coomassie system using RanGAP-SUMO as the substrate, thus avoiding the use of expensive fluorescent dyes or the error-prone fluorescent reporter. Its reproducibility and reliability were also evaluated in this report.
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Xie
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Zhongli
authors.initials: Z
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Jianchen
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Lie
authors.initials: L
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Yaxue
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: hczhou@sjtu.edu.cn.
authors.forename: Huchen
authors.initials: H
authors.lastname: Zhou
mesh_terms: D001549:Benzamides / Q000032:analysis; D004396:Coloring Agents; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics; D057075:Enzyme Assays / Q000379:methods*; D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D011480:Protease Inhibitors / Q000032:analysis*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D012394:Rosaniline Dyes
chemical_list: D001549:Benzamides; D004396:Coloring Agents; D020690:GTPase-Activating Proteins; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; C404512:SENP1 protein, human; D003546:Cysteine Endopeptidases; C501833:SENP2 protein, human
keywords: Assay; DeSUMOlyation; Inhibitor; SENP; SUMO