FRET-based in vitro assays for the analysis of SUMO protease activities

    Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

    17 Citations (Scopus)

    Abstract

    In humans cells three SUMO paralogues (SUMO-1, SUMO-2 and SUMO-3) and six SUMO specific proteases (SENP1-SENP3 and SENP5-SENP7) are expressed. Together the SUMO proteases perform three distinct functions. They: (1) process the immature pro-SUMO proteins into the active forms, (2) remove SUMO molecules conjugated to protein targets, and (3) depolymerise SUMO conjugated within polymeric chains. By regulating these processes the SENPs play a crucial role in regulating the sumoylation state of target proteins in cells, and therefore are academically and pharmacologically interesting enzymes. Gel-based techniques for SENP analysis are well established and can be used for many applications, but their laborious methodology makes them cumbersome tools for kinetic analysis or inhibitor screening. Therefore in vitro FRET-based assays have been developed to test the three major functions of the SENPs. These use fluorescent protein fusions of the SUMOs, and together facilitate high-throughput, real-time analysis of the three major SUMO protease activities.
    Original languageEnglish
    Title of host publicationSUMO protocols
    EditorsHelle D. Ulrich
    PublisherHumana Press
    Pages253-268
    Number of pages16
    Volume6
    ISBN (Electronic)9781597455664
    ISBN (Print)9781934115800
    DOIs
    Publication statusPublished - 2009

    Publication series

    NameMethods in Molecular Biology
    PublisherHumana Press
    Volume497
    ISSN (Print)1064-3745

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