Eisenhardt, N., Chaugule, V. K. and Pichler, A. (2016) A fluorescent in vitro assay to investigate paralog-specific SUMO conjugation. In: Rodriguez, M. (ed.) SUMO. Series: Methods in molecular biology, 1475 (1475). Humana Press, pp. 67-78. ISBN 9781493963560 (doi: 10.1007/978-1-4939-6358-4_5)
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Abstract
Protein modification with the small ubiquitin-related modifier SUMO is a potent regulatory mechanism implicated in a variety of biological pathways. In vitro sumoylation reactions have emerged as a versatile tool to identify and characterize novel SUMO enzymes as well as their substrates. Here, we present detailed protocols for the purification and fluorescent labeling of mammalian SUMO paralogs for their application in sumoylation assays. These assays provide a fast readout for in vitro SUMO chain formation activity of E3 ligases in a paralog-specific manner. Finally, we critically analyze the application of fluorescent SUMO proteins to study substrate modification in vitro revealing also the drawbacks of the system.
| Item Type: | Book Sections |
|---|---|
| Additional Information: | Online ISBN: 9781493963584. |
| Status: | Published |
| Glasgow Author(s) Enlighten ID: | Chaugule, Dr Viduth |
| Authors: | Eisenhardt, N., Chaugule, V. K., and Pichler, A. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Publisher: | Humana Press |
| ISSN: | 1064-3745 |
| ISBN: | 9781493963560 |
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