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

Mikako Hirohama, Arnout R D Voet, Takeaki Ozawa, Hisato Saitoh, Youichi Nakao, Kam Y J Zhang, Akihiro Ito, Minoru Yoshida

    Research output: Contribution to journalArticle

    7 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)92-94
    Number of pages3
    JournalAnalytical Biochemistry
    Volume448
    Issue number1
    DOIs
    Publication statusPublished - 2014 Mar 1

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    Keywords

    • Assay
    • Protein-protein interactions
    • SIM
    • SUMOylation

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Molecular Biology
    • Cell Biology

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