Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system.
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.
['Hirohama|Mikako|M|', 'Voet|Arnout R D|AR|', 'Ozawa|Takeaki|T|', 'Saitoh|Hisato|H|', 'Nakao|Yoichi|Y|', 'Zhang|Kam Y J|KY|', 'Ito|Akihiro|A|', 'Yoshida|Minoru|M|']
D000595:Amino Acid Sequence D002352:Carrier Proteins D004926:Escherichia coli D005456:Fluorescent Dyes D017930:Genes, Reporter D006801:Humans D008027:Light D008156:Luciferases D054730:Protein Interaction Domains and Motifs D017434:Protein Structure, Tertiary D011993:Recombinant Fusion Proteins D025842:SUMO-1 Protein D013050:Spectrometry, Fluorescence
Assay SIM SUMOylation Protein–protein interactions