Purification and identification of endogenous polySUMO conjugates.
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.
Wellcome Trust Centre for Gene Regulation and Expression, Dow Street, Dundee DD1 5EH, UK.
Roland
R
Bruderer
Michael H
MH
Tatham
Anna
A
Plechanovova
Ivan
I
Matic
Amit K
AK
Garg
Ronald T
RT
Hay
D000595:Amino Acid Sequence; D006367:HeLa Cells; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D020543:Proteome / Q000302:isolation & purification* / Q000378:metabolism; D013312:Stress, Physiological; D013347:Subcellular Fractions / Q000378:metabolism; D058207:Sumoylation; D014157:Transcription Factors / Q000378:metabolism*
D009687:Nuclear Proteins; D020543:Proteome; C114070:RNF4 protein, human; D014157:Transcription Factors
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