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

    Fingerprint

    Ubiquitin
    Luciferases
    Assays
    Sumoylation
    Proteins
    In Vitro Techniques
    Screening
    Throughput

    Keywords

    • Assay
    • Protein-protein interactions
    • SIM
    • SUMOylation

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Molecular Biology
    • Cell Biology

    Cite this

    Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system. / Hirohama, Mikako; Voet, Arnout R D; Ozawa, Takeaki; Saitoh, Hisato; Nakao, Youichi; Zhang, Kam Y J; Ito, Akihiro; Yoshida, Minoru.

    In: Analytical Biochemistry, Vol. 448, No. 1, 01.03.2014, p. 92-94.

    Research output: Contribution to journalArticle

    Hirohama, Mikako ; Voet, Arnout R D ; Ozawa, Takeaki ; Saitoh, Hisato ; Nakao, Youichi ; Zhang, Kam Y J ; Ito, Akihiro ; Yoshida, Minoru. / Assay methods for small ubiquitin-like modifier (SUMO)-SUMO-interacting motif (SIM) interactions in vivo and in vitro using a split-luciferase complementation system. In: Analytical Biochemistry. 2014 ; Vol. 448, No. 1. pp. 92-94.
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    AU - Ozawa, Takeaki

    AU - Saitoh, Hisato

    AU - Nakao, Youichi

    AU - Zhang, Kam Y J

    AU - Ito, Akihiro

    AU - Yoshida, Minoru

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