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Free keywords:
Amino Acid Sequence
HeLa Cells
Hot Temperature
Humans
Molecular Sequence Data
Nuclear Proteins/*metabolism
Protein Binding
Protein Interaction Domains and Motifs
Proteome/*isolation & purification/metabolism
Stress, Physiological
Subcellular Fractions/metabolism
Sumoylation
Transcription Factors/*metabolism
Abstract:
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.
