In vivo identification of human small ubiquitin-like modifier polymerization sites by high accuracy mass spectrometry and an in vitro to in vivo strategy

Ivan Matic, Martijn van Hagen, Joost Schimmel, Boris Macek, Stephen C. Ogg, Michael H. Tatham, Ronald T. Hay, Angus I. Lamond, Matthias Mann (Lead / Corresponding author), Alfred C. O. Vertegaal (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    198 Citations (Scopus)

    Abstract

    The length and precise linkage of polyubiquitin chains is important for their biological activity. Although other ubiquitin-like proteins have the potential to form polymeric chains their identification in vivo is challenging and their functional role is unclear. Vertebrates express three small ubiquitin-like modifiers, SUMO-1, SUMO-2, and SUMO-3. Mature SUMO-2 and SUMO-3 are nearly identical and contain an internal consensus site for sumoylation that is missing in SUMO-1. Combining state-of-the-art mass spectrometry with an "in vitro to in vivo" strategy for post-translational modifications, we provide direct evidence that SUMO-1, SUMO-2, and SUMO-3 form mixed chains in cells via the internal consensus sites for sumoylation in SUMO-2 and SUMO-3. In vitro, the chain length of SUMO polymers could be influenced by changing the relative amounts of SUMO-1 and SUMO-2. The developed methodology is generic and can be adapted for the identification of other sumoylation sites in complex samples.

    Original languageEnglish
    Pages (from-to)132-144
    Number of pages13
    JournalMolecular & Cellular Proteomics
    Volume7
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2008

    Keywords

    • SUMO-2 TARGET PROTEINS
    • QUANTITATIVE PROTEOMICS
    • SUMOYLATION SITES
    • CELL-CYCLE
    • E3 LIGASE
    • PATHWAY
    • COMPLEX
    • CHAINS
    • YEAST
    • UBC9

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