Abstract
Small ubiquitin-related modifier (SUMO) is an ubiquitin-like protein that is covalently attached to a variety of target proteins. Unlike ubiquitination, sumoylation does not target proteins for proteolytic breakdown, but is instead involved in regulating multiple protein functional properties including protein−protein interactions and subcellular targeting, to name a few. Protein sumoylation has been particularly well characterized as a regulator of many nuclear processes as well as nuclear structure, making the characterization of this modification vital for understanding nuclear structure and function. Consequently, there has been intense interest in identifying new proteins that are targets of this modification and determining what role it plays in regulating their functions. This chapter presents methodologies for determining whether a particular protein is a substrate of sumoylation, and for identifying the lysine residue(s) where the modification occurs.
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Acknowledgments
We are very grateful to Mike Matunis (Johns Hopkins), Ron Hay (University of Dundee), and Chris Lima (Sloan Kettering Institute) for providing constructs and reagents.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Sarge, K.D., Park-Sarge, OK. (2009). Detection of Proteins Sumoylated In Vivo and In Vitro. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_17
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DOI: https://doi.org/10.1007/978-1-60327-378-7_17
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