Proteome-wide identification of SUMO2 modification sites

Triin Tammsalu, Ivan Matic, Ellis G. Jaffray, Adel F. M. Ibrahim, Michael H. Tatham, Ronald T. Hay (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    107 Citations (Scopus)

    Abstract

    Posttranslational modification with small ubiquitin-like modifiers (SUMOs) alters the function of proteins involved in diverse cellular processes. SUMO-specific enzymes conjugate SUMOs to lysine residues in target proteins. Although proteomic studies have identified hundreds of sumoylated substrates, methods to identify the modified lysines on a proteomic scale are lacking. We developed a method that enabled proteome-wide identification of sumoylated lysines that involves the expression of polyhistidine (6His)-tagged SUMO2 with Thr90 mutated to Lys. Endoproteinase cleavage with Lys-C of 6His-SUMO2T90K-modified proteins from human cell lysates produced a diGly remnant on SUMO2T90K-conjugated lysines, enabling immunoprecipitation of SUMO2T90K-modified peptides and producing a unique mass-to-charge signature. Mass spectrometry analysis of SUMO-enriched peptides revealed more than 1000 sumoylated lysines in 539 proteins, including many functionally related proteins involved in cell cycle, transcription, and DNA repair. Not only can this strategy be used to study the dynamics of sumoylation and other potentially similar posttranslational modifications, but also, these data provide an unprecedented resource for future research on the role of sumoylation in cellular physiology and disease.

    Original languageEnglish
    Article numberrs2
    Number of pages10
    JournalScience Signaling
    Volume7
    Issue number323
    DOIs
    Publication statusPublished - 29 Apr 2014

    Fingerprint

    Proteome
    Lysine
    Ubiquitin
    Sumoylation
    Post Translational Protein Processing
    Proteins
    Proteomics
    Cells
    Peptides
    Physiology
    Transcription
    Immunoprecipitation
    DNA Repair
    Mass spectrometry
    Mass Spectrometry
    Cell Cycle
    Repair
    DNA
    Substrates
    Enzymes

    Cite this

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    title = "Proteome-wide identification of SUMO2 modification sites",
    abstract = "Posttranslational modification with small ubiquitin-like modifiers (SUMOs) alters the function of proteins involved in diverse cellular processes. SUMO-specific enzymes conjugate SUMOs to lysine residues in target proteins. Although proteomic studies have identified hundreds of sumoylated substrates, methods to identify the modified lysines on a proteomic scale are lacking. We developed a method that enabled proteome-wide identification of sumoylated lysines that involves the expression of polyhistidine (6His)-tagged SUMO2 with Thr90 mutated to Lys. Endoproteinase cleavage with Lys-C of 6His-SUMO2T90K-modified proteins from human cell lysates produced a diGly remnant on SUMO2T90K-conjugated lysines, enabling immunoprecipitation of SUMO2T90K-modified peptides and producing a unique mass-to-charge signature. Mass spectrometry analysis of SUMO-enriched peptides revealed more than 1000 sumoylated lysines in 539 proteins, including many functionally related proteins involved in cell cycle, transcription, and DNA repair. Not only can this strategy be used to study the dynamics of sumoylation and other potentially similar posttranslational modifications, but also, these data provide an unprecedented resource for future research on the role of sumoylation in cellular physiology and disease.",
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    Proteome-wide identification of SUMO2 modification sites. / Tammsalu, Triin; Matic, Ivan; Jaffray, Ellis G.; Ibrahim, Adel F. M.; Tatham, Michael H.; Hay, Ronald T. (Lead / Corresponding author).

    In: Science Signaling, Vol. 7, No. 323, rs2, 29.04.2014.

    Research output: Contribution to journalArticle

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    AU - Matic, Ivan

    AU - Jaffray, Ellis G.

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    AU - Tatham, Michael H.

    AU - Hay, Ronald T.

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