title: Site-specific mapping and quantification of protein S-sulphenylation in cells.
abstract: Cysteine S-sulphenylation provides redox regulation of protein functions, but the global cellular impact of this transient post-translational modification remains unexplored. We describe a chemoproteomic workflow to map and quantify over 1,000 S-sulphenylation sites on more than 700 proteins in intact cells. Quantitative analysis of human cells stimulated with hydrogen peroxide or epidermal growth factor measured hundreds of site selective redox changes. Different cysteines in the same proteins displayed dramatic differences in susceptibility to S-sulphenylation. Newly discovered S-sulphenylations provided mechanistic support for proposed cysteine redox reactions and suggested novel redox mechanisms, including S-sulphenyl-mediated redox regulation of the transcription factor HIF1A by SIRT6. S-sulphenylation is favored at solvent-exposed protein surfaces and is associated with sequence motifs that are distinct from those for other thiol modifications. S-sulphenylations affect regulators of phosphorylation, acetylation and ubiquitylation, which suggest regulatory crosstalk between redox control and signalling pathways.
authors.affiliation: 1] Departmemt of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6350, USA [2] Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6350, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, Florida 33458, USA.
authors.forename: Vinayak
authors.initials: V
authors.lastname: Gupta
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Jupiter, Florida 33458, USA.
authors.forename: Kate S
authors.initials: KS
authors.lastname: Carroll
authors.affiliation: 1] Departmemt of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6350, USA [2] Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6350, USA.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Liebler
mesh_terms: D000107:Acetylation; D045744:Cell Line, Tumor; D003545:Cysteine / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D058977:Molecular Sequence Annotation; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D037761:Sirtuins / Q000235:genetics / Q000378:metabolism*; D013434:Sulfenic Acids / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D013434:Sulfenic Acids; D004815:Epidermal Growth Factor; D006861:Hydrogen Peroxide; C516592:SIRT6 protein, human; D037761:Sirtuins; D003545:Cysteine