title: Identification of Substrates of Protein-Group SUMOylation.
abstract: Protein modification by conjugation to the ubiquitin-related protein SUMO (SUMOylation) regulates numerous cellular functions and is reversible. However, unlike typical posttranslational modifications, SUMOylation often targets and regulates proteins of functionally and physically linked protein groups, rather than individual proteins. Functional studies of protein-group SUMOylation are thus particularly challenging, as they require the identification of ideally all members of a modified protein group. Here, we describe mass spectrometric approaches to detect SUMOylated protein groups in Saccharomyces cerevisiae, yet the protocols can be readily adapted for studies of SUMOylation in mammalian cells.
authors.affiliation: Department of Molecular Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany. ivan.psakhye@ifom.eu.|IFOM, the FIRC Institute of Molecular Oncology, Via Adamello 16, 20139, Milan, Italy. ivan.psakhye@ifom.eu.
authors.forename: Ivan
authors.initials: I
authors.lastname: Psakhye
authors.affiliation: Department of Molecular Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany. jentsch@biochem.mpg.de.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jentsch
mesh_terms: D002846:Chromatography, Affinity; D007553:Isotope Labeling; D013058:Mass Spectrometry; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D058207:Sumoylation
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D025841:Small Ubiquitin-Related Modifier Proteins
keywords: Affinity purification; Mass spectrometry; Protein complex; Protein-group SUMOylation; Quantitative proteomics; SUMO; Stable isotope labeling by amino acids in cell culture (SILAC)