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  Analysis of Sumoylation

Breucker, J., & Pichler, A. (2019). Analysis of Sumoylation. Methods in Molecular Biology, 1934, 223-233. doi:10.1007/978-1-4939-9055-9_14.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0004-A636-4 Version Permalink: http://hdl.handle.net/21.11116/0000-0004-DAAD-4
Genre: Journal Article

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 Creators:
Breucker , Jan1, Author
Pichler, Andrea1, Author              
Affiliations:
1Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, 79108 Freiburg, DE, ou_2243640              

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Free keywords: SUMO, SUMO1, SUMO2/3, MBD1, E1-activating enzyme, E2-conjugating enzyme, E3-ligase, RanBP2ΔFG, RanBP2 IR1+M, PIAS1
 Abstract: Protein regulation by reversible attachment of SUMO (small ubiquitin-related modifier) plays an important role in several cellular processes such as transcriptional regulation, nucleo-cytoplasmic transport, cell-cycle progression, meiosis, and DNA repair. However, most sumoylated proteins are of marginal abundance at steady state levels, which is due to strict regulation and/or rapid turnover of modification and de-modification. Consequently, analysis of protein sumoylation in vivo is very challenging. Nonetheless, a novel method was established that allows detection of sumoylated proteins at endogenous levels from vertebrate cells and tissues. This approach involves the enrichment of sumoylated proteins by immunoprecipitation followed by peptide elution. After endogenous substrate sumoylation is verified, addressing its functional consequences is the next logical step. This requires SUMO site mapping that benefits from larger quantities of modified protein. Here, we shortly describe strategies to achieve efficient in vitro sumoylation of many substrates.

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Language(s): eng - English
 Dates: 2019
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1007/978-1-4939-9055-9_14
 Degree: -

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Title: Methods in Molecular Biology
  Other : Methods Mol. Biol.
Source Genre: Journal
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Affiliations:
Publ. Info: Clifton, N.J. : Humana Press
Pages: - Volume / Issue: 1934 Sequence Number: - Start / End Page: 223 - 233 Identifier: ISSN: 1064-3745
CoNE: https://pure.mpg.de/cone/journals/resource/954927725544