Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system

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• 作者：Mikako Hirohama ; Arnout R.D. Voet ; Takeaki Ozawa ; Hisato Saitoh ; Yoichi Nakao ; Kam Y.J. Zhang ; Akihiro Ito ; Minoru Yoshida
• 关键词：Assay ; SIM ; SUMOylation ; Protein&ndash ; protein interactions
• 刊名：Analytical Biochemistry
• 出版年：1 March, 2014
• 年：2014
• 卷：448
• 期：Complete
• 页码：92-94
• 全文大小：386 K

文摘

SUMOylation is a posttranslational process that attaches a small ubiquitin-like modifier (SUMO) to its target proteins covalently. SUMOylation controls multiple cellular processes through the recognition of SUMO by a SUMO-interacting motif (SIM). In this study, we developed assay systems for detecting noncovalent interactions between SUMO and SIM in cells using split-luciferase complementation. We applied a version of this assay to the detection of in vitro SUMO-SIM interactions using a bacterial expression system. These novel assays enable screening of inhibitors of SUMO-dependent protein-protein interactions, either in vivo or in vitro, in a high-throughput manner.