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Proteomics Strategies to Identify SUMO Targets and Acceptor Sites: A Survey of RNA-Binding Proteins SUMOylation

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Abstract

SUMOylation is a protein posttranslational modification that participates in the regulation of numerous biological processes within the cells. Small ubiquitin-like modifier (SUMO) proteins are members of the ubiquitin-like protein family and, similarly to ubiquitin, are covalently linked to a lysine residue on a target protein via a multi-enzymatic cascade. To assess the specific mechanism triggered by SUMOylation, the identification of SUMO protein substrates and of the precise acceptor site to which SUMO is bound is of critical relevance. Despite hundreds of mammalian proteins have been described as targets of SUMOylation, the identification of the precise acceptor sites still represents an important analytical challenge because of the relatively low stoichiometry in vivo and the highly dynamic nature of this modification. Moreover, mass spectrometry-based identification of SUMOylated sites is hampered by the large peptide remnant of SUMO proteins that are left on the modified lysine residue upon tryptic digestion. The present review provides a survey of the strategies that have been exploited in order to enrich, purify and identify SUMOylation substrates and acceptor sites in human cells on a large-scale format. The success of the presented strategies helped to unravel the numerous activities of this modification, as it was shown by the exemplary case of the RNA-binding protein family, whose SUMOylation is here reviewed.

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Authors and Affiliations

  1. Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, 20126, Milan, Italy

    Giuseppe Filosa & Silvia M. L. Barabino

  2. IFOM, FIRC Institute of Molecular Oncology, via Adamello 16, 20139, Milan, Italy

    Giuseppe Filosa & Angela Bachi

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  1. Giuseppe Filosa
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  2. Silvia M. L. Barabino
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  3. Angela Bachi
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Correspondence to Angela Bachi.

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Filosa, G., Barabino, S.M.L. & Bachi, A. Proteomics Strategies to Identify SUMO Targets and Acceptor Sites: A Survey of RNA-Binding Proteins SUMOylation. Neuromol Med 15, 661–676 (2013). https://doi.org/10.1007/s12017-013-8256-8

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  • DOI: https://doi.org/10.1007/s12017-013-8256-8

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