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SUMO pp 109–121Cite as

Using Biotinylated SUMO-Traps to Analyze SUMOylated Proteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1475))

Abstract

SUMO-interacting motifs (SIMs) recognize SUMOylated proteins with high specificity allowing to connect SUMO-modified proteins. Multiple SIMs fused to distinct tags have been used to increase their affinity and generate more efficient purification tools. Enrichment of SUMOylated proteins using SIMs arranged in tandem (SUMO-traps) facilitates the identification and characterization of protein targets in vitro and in vivo. Here a protocol to produce biotinylated SUMO-traps (bioSUBEs) to capture SUMO chains and typical SUMOylated proteins such as p53 or IkBα is presented. Biotinylated SUMO-traps represent an alternative to reduce the background associated to bigger tags, e.g., during mass spectrometry analysis. Consequently, bioSUBEs are alternative tools to characterize endogenous SUMO targets.

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Acknowledgements

UCMB at Inbiomed was supported by the “Obra Social KUTXA” and the Diputación Foral de Gipuzkoa. The author(s) would like to acknowledge networking support by the PROTEOSTASIS action BM1307, supported by COST (European Cooperation in Science and Technology). RB acknowledges the support of the Spanish MINECO (BFU2014-52282-P).

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Authors and Affiliations

  1. Ubiquitylation and Cancer MolecularBiology (UMCB) Laboratory, Inbiomed, 20009, San Sebastian-Donostia, Gipuzkoa, Spain

    Valérie Lang & Elisa Da Silva-Ferrada

  2. Group of Ubiquitin-Dependent Proteolysis and Intracellular Communication (G(u)ic), Institute of Biomedical Imaging and Life Sciences (IBILI), University of Coimbra, 3000-548, Coimbra, Portugal

    Elisa Da Silva-Ferrada

  3. CIC bioGUNE, Bizkaia Technology Park, 48160, Derio, Bizkaia, Spain

    Rosa Barrio & James D. Sutherland

  4. INBIOMED, San Sebastian, Spain

    Manuel S. Rodriguez

Authors
  1. Valérie Lang
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  2. Elisa Da Silva-Ferrada
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  3. Rosa Barrio
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  4. James D. Sutherland
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  5. Manuel S. Rodriguez
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Corresponding author

Correspondence to Manuel S. Rodriguez .

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Editors and Affiliations

  1. INBIOMED, San Sebastian, Spain

    Manuel S. Rodriguez

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Lang, V., Da Silva-Ferrada, E., Barrio, R., Sutherland, J.D., Rodriguez, M.S. (2016). Using Biotinylated SUMO-Traps to Analyze SUMOylated Proteins. In: Rodriguez, M. (eds) SUMO. Methods in Molecular Biology, vol 1475. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6358-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6358-4_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6356-0

  • Online ISBN: 978-1-4939-6358-4

  • eBook Packages: Springer Protocols

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