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Purification and identification of endogenous polySUMO conjugates
Author(s) -
Bruderer Roland,
Tatham Michael H,
Plechanovova Anna,
Matic Ivan,
Garg Amit K,
Hay Ronald T
Publication year - 2011
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/embor.2010.206
Subject(s) - ubiquitin , tandem affinity purification , endogeny , affinity chromatography , biochemistry , biology , genome , conjugate , chemistry , computational biology , enzyme , microbiology and biotechnology , gene , mathematical analysis , mathematics
The small ubiquitin‐like modifier (SUMO) can undergo self‐modification to form polymeric chains that have been implicated in cellular processes such as meiosis, genome maintenance and stress response. Investigations into the biological role of polymeric chains have been hampered by the absence of a protocol for the purification of proteins linked to SUMO chains. In this paper, we describe a rapid affinity purification procedure for the isolation of endogenous polySUMO‐modified species that generates highly purified material suitable for individual protein studies and proteomic analysis. We use this approach to identify more than 300 putative polySUMO conjugates from cultured eukaryotic cells.