Purification and identification of endogenous polySUMO conjugates

• Published in: EMBO reports Vol. 12; no. 2; pp. 142 - 148
• Main Authors: Bruderer, Roland, Tatham, Michael H, Plechanovova, Anna, Matic, Ivan, Garg, Amit K, Hay, Ronald T
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-02-2011; Nature Publishing Group UK; Blackwell Publishing Ltd; Nature Publishing Group
• Subjects: affinity purification; Amino Acid Sequence; Cellular biology; EMBO31; HeLa Cells; Hot Temperature; Humans; mass spectrometry; Molecular biology; Molecular Sequence Data; Nuclear Proteins - metabolism; polySUMO; Protein Binding; Protein Interaction Domains and Motifs; Proteins; Proteome - isolation & purification; Proteome - metabolism; Proteomics; Scientific Report; Scientific Reports; Stress, Physiological; Subcellular Fractions - metabolism; SUMO; SUMO substrates; Sumoylation; Transcription Factors - metabolism; mass spectrometry; polySUMO; SUMO; affinity purification; SUMO substrates
• ISSN: 1469-221X; 1469-3178
• DOI: 10.1038/embor.2010.206

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.
The authors describe a rapid affinity purification procedure for the isolation of endogenous polySUMO modified proteins, which provides highly purified material suitable for individual protein studies as well as proteomic analysis.