Detection and quantitation of SUMO chains by mass spectrometry

Ivan Matic, Ronald T. Hay

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

    7 Citations (Scopus)

    Abstract

    The small ubiquitin-like modifiers (SUMOs) alter the function of cellular proteins by covalent attachment to lysine side-chains. SUMOs can target themselves for modification so generating SUMO polymers, the functions of which are beginning to be unraveled. The identification and quantitation of SUMO chains is essential for the functional investigation of SUMO polymerization. Classical techniques, such as site-directed mutagenesis and western blotting, are indirect and often inconclusive methods for the study of SUMO polymers. On the contrary, direct detection is possible with mass spectrometry (MS) by the identification of the SUMO-SUMO branched peptide remnant after proteolytic digestion. In this chapter, we describe a straightforward workflow that incorporates a modified database to efficiently detect SUMO polymers from simple and complex protein samples. In combination with stable isotope labeling by amino acids in cell culture (SILAC), this proteomic strategy allows accurate relative quantitation of SUMO polymers from different biological samples.
    Original languageEnglish
    Title of host publicationUbiquitin family modifiers and the proteasome
    Subtitle of host publicationreviews and protocols
    EditorsR. Jürgen Dohmen, Martin Scheffner
    PublisherHumana Press
    Pages239-247
    Number of pages9
    ISBN (Electronic)9781617794742
    ISBN (Print)9781617794735
    DOIs
    Publication statusPublished - 1 Jan 2012

    Publication series

    NameMethods in molecular biology
    PublisherHumana Press
    Volume832
    ISSN (Print)1064-3745

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  • Cite this

    Matic, I., & Hay, R. T. (2012). Detection and quantitation of SUMO chains by mass spectrometry. In R. J. Dohmen, & M. Scheffner (Eds.), Ubiquitin family modifiers and the proteasome: reviews and protocols (pp. 239-247). (Methods in molecular biology; Vol. 832). Humana Press. https://doi.org/10.1007/978-1-61779-474-2_17