MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v01/MSV000078777/primaryOK2000000ProteomicsDaniel C. LieblerQ ExactiveHomo Sapiens (ncbitaxon:9606)https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=e3d5ee78d54a44e7b0f7febfdb479292MassIVE54MOD:00397 - "A protein modification that is produced by reaction with iodoacetamide, usually replacement of a reactive hydrogen with a methylcarboxamido group."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.Site-specific mapping and quantification of protein S-sulphenylation in cells.Yang Jing J, Gupta Vinayak V, Carroll Kate S KS, Liebler Daniel C DCliquid chromatography tandem mass spectroscopy, Archive, Click Chemical, Activity, Chemical Reaction, Laboratory, Reagent, Chemical Technique, FBN, Gene, high weight, Click Chemical Reaction, Visible Light, Dmel_CG9403, Click Chemical Reactions, Strepavidin, heavy chain disease, Medical Specialties, LC-MS-MS, Ly113, peptide, Speciality, Polypeptides, Techniques, Medical Specialities, peptido, Specialities, ECTOL1, LC-MSMS, hnu, heavy, Line, Gene Products, 1.28, anon-WO0140519.173, Research Activity, Vitamin K Dependent Protein S, Medical, Laboratory Research, Callisto, Reagents and Indicators, Technique, foton, WMS, Priorities, BcDNA:LD21358, l(2)22F3, Vitamin K Dependent, Insurance Medicine, Io, Vitamin K-Dependent Protein S, LCMSMS, reactivo, peptides, jupiter, CG9397, Research, DmelCG31363, Lichtquant, beta-Trypsin, Visible, Indicators, OCTD, CG31363, TNFSF14, drugs, 2-iodo-., medicine, Reaction, CT9093, Click Chemical Technique, Medicine, photon, site, peptidos, Medicines, Development and Research, TR2, Insurance, Research Priority, Reagents, CG17238, GPHYSD2, Jing, reagents, gamma, SGS, LC-MS2, Specialties, Radiation, Papers, Click, Medical Specialty, UNQ391/PRO726, Cofactor, Proteins, LC-MS/MS, Cell Lines, Research Priorities, Chemical Reactions, CG9403, Light, clone 1.76, CD258, Cell, Peptide, Ganymede, l(2)01094, ACMICD, Chemical Techniques, polypeptide, Medical Speciality, BcDNA:LD43690, LC/MS/MS, Priority, Cofactor Protein S, BEST:LP05168, LIGHT, HCD, Protein, Research Activities, Specialty, BcDNA:GH06373, Chemistry, MFS1, University, reactif, region, WMS2, Lines, Research and Development, Visible Radiations, Radiations, Visible Radiation, PRSS, Reactions, HVEML, HVEM-L, Photoradiation, Tripcellim, anon-EST:Liang-1.76, reagent, MASS, DmelCG9397, Europa, beta Trypsin, LTg, Activities, Protein Gene Products, Click Chemical Techniques, Insurance Medicines, Gene Proteins, Trypure, Click Chemistries, light quantum, BcDNA:RH53211, Photoradiations, label, liquid chromatography-tandem mass spectroscopy, Acetamide, Indicator, SSKS, liquid chromatography tandem mass spectrometry, Peptid, Polypeptide, Chemistries, Protein S, Vitamin K-Dependent, CG18762, CG18763Post Translational Amino Acid Modification, T-cell leukemia, posttranslational modification, E 920, methionine aminopeptidase activity, determination, H2O2, Compounds, AGL4, Oxydol, TGF-alpha receptor binding, protein, Protein Processing, dihydrogen dioxide, Solvent, phosphorylation, Social Controls, prevention, Thiol, protein polypeptide chains, Human Urinary Gastric Inhibitor, hif-1alpha, RNA polymerase II distal enhancer sequence-specific DNA binding transcription factor activity, 2, 2-amino-3-mercaptopropanoic acid, Cystein, protein aggregate, Work Flow, prevention and control, Formal Social Controls, peptidase M activity, imprinted and ancient gene protein, adenomas, Post-Translational Protein, C, rabGAPLP, perhydrol, posttranslational amino acid modification, L-methionine aminopeptidase activity, Ubiquitylation, Migrant Worker, Man (Taxonomy), Hydroperoxide, reference sample, Post-Translational, cisteina, Moods, Nonmigrant, F10N7_150, RabGAP-5, proteins, Growth Factor-Urogastrone, Posttranslational Protein Processing, Transient, RNA polymerase II core promoter proximal region sequence-specific DNA binding transcription factor activity, Social, preventive measures, transforming growth factor alpha receptor binding, HIF-1alpha, RUSC3, signaling process, HIF1a, Squatters, AGAMOUS-like 4, ARNT-interacting protein, zinc ion regulated proximal promoter sequence-specific DNA binding, sequence-specific transcription regulatory region DNA binding RNA polymerase II transcription factor recruiting transcription factor activity, Half Cystine, TRANSCRIPTION FACTOR, single organism signaling, RNA polymerase II distal enhancer sequence-specific binding, distinct from, RNA polymerase II core promoter proximal region sequence-specific binding, Post-Translational Modification, Thiols, Mercaptans, preventive therapy, Zinc Cysteinate, Modern, Modifications, Mercapto Compounds, acetylation, MYH-associated polyposis, autosomal recessive familial adenomatous polyposis, Maps, metal ion regulated sequence-specific DNA binding RNA polymerase II transcription factor activity, HIF-1A, Cys, Workflows, Post Translational Modifications, Perhydrol, Migrant, beta Urogastrone, Workers, sequence-specific distal enhancer binding RNA polymerase II transcription factor activity, (2R)-2-amino-3-sulfanylpropanoic acid, Urogastrone, Migrants and Transients, region, autosomal recessive, F14P3.4, RUTBC3, Superoxol, Factors, PTM, Hydrogen peroxide (H2O2), Processing, Control, F14P3_4, Epidermal growth factor, L-Cysteine, Controls, exposed, Transcription factor, human, L-Cystein, RABGAP5, Amino Acid Modification, post-translational modification, BHLHE78, multiple colorectal, copper ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, PASD8, Migrants, Posttranslational Modifications, E430016J11Rik, Regulation, TGFalpha receptor binding, metal ion regulated sequence-specific DNA binding, Posttranslational Modification, Work Flows, humans, metal ion regulated proximal promoter sequence-specific DNA binding, Regulations, HOOH, EGF receptor binding, biological signaling, Cysteine Hydrochloride, human being, Post-Translational Protein Modifications, Sulfhydryl, number, L-Zystein, Gene, SEPALLATA 2, protein-containing complex, Hydrogen Peroxide (H2O2), Ximpact, Transcription Factor, presence, AI043036, Migrant Workers, AA959795, Posttranslational, Human, beta-Urogastrone, zinc ion regulated core promoter proximal region sequence-specific DNA binding, MOP1, polypeptide chain, Homo sapiens, SIR2L6, Gene Products, Post Translational Modification, Mood, Post-Translational Modifications, Growth Factor, Half-Cystine, Man, familial adenomatous polyposis, epidermal growth factor, Post Translational, MAP, Modification, Transcription, transforming growth factor alpha receptor ligand, epidermal growth factor receptor ligand, L Cysteine, HIF1, E-920, familial adenomatous polyposis 2, Transients, EGFR binding, Acetylations, Nonmigrants, Peroxide, Hcys, posttranslational protein modification, Worker, Sir2l6, 2810449N18Rik, Protein Modifications, autosomal recessive multiple colorectal adenomas, Squatter, Post-Translational Amino Acid Modification, Epidermal Growth Factor-Urogastrone, extruding from, CYSTEINE, Posttranslational Amino Acid Modification, RNA polymerase II proximal promoter sequence-specific DNA binding, [OH(OH)], copper ion regulated proximal promoter sequence-specific DNA binding, EGF receptor ligand, Posttranslational Protein, FREE CYSTEINE, site, (2R)-2-amino-3-mercaptopropanoic acid, Controlled, hif1a, signalling., Controlling, MUTYH-Associated Polyposis, pro-epidermal growth factor, Formal Social Control, protein complex, exits through, Affects, Proteins, Phosphorylations, Post-Translational Protein Modification, zinc ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, homeobox 1, Factor, hif1alpha, impact-a, Cell, transcription factor activity, count in organism, Mercaptan, transforming growth factor alpha, native protein, natural protein, Social Control, xhif1a, Protein, chemical analysis, Nomad, RNA polymerase II transcription factor activity, sequence, IMPACT, imprinted and ancient gene protein homolog, Protein Modification, metal ion regulated core promoter proximal region sequence-specific DNA binding RNA polymerase II transcription factor activity, KNOTTED1-like homeobox gene 5, Zystein, susceptibility, Hydrogen, Sulfhydryl Compound, 2-amino-3-sulfanylpropanoic acid, Post Translational Protein Processing, HIF1-ALPHA, prophylaxis, L-2-Amino-3-mercaptopropionic acid, (R)-2-amino-3-mercaptopropanoic acid, MYH-Associated Polyposis, metal ion regulated core promoter proximal region sequence-specific binding, copper ion regulated core promoter proximal region sequence-specific binding, 2-Amino-3-mercaptopropionic acid, F10N7.150, primary structure of sequence macromolecule, post-translational amino acid modification, Protein Gene Products, Gene Proteins, HIF1alpha, Post Translational Protein Modification, EGF, Epidermal, signalling process, Compound, Mercapto, control, Isolation of Nuclei TAgged in specific Cell Types, Nomads, Modern Man, Post-Translational Protein Processing, E920, MAP syndrome, regulation, bHLHe78, assay, RWDD5, FAP2, colorectal adenomatous polyposissite, Cell., Vitamin K Dependent, Vitamin K-Dependent Protein S, Cofactor Protein S, Vitamin K Dependent Protein S, Protein S, region, Vitamin K-Dependent, Cofactorsite, Cell., Vitamin K Dependent, Vitamin K-Dependent Protein S, Cofactor Protein S, Vitamin K Dependent Protein S, Protein S, region, Vitamin K-Dependent, Cofactor0trueYang: Site-specific mapping and quantification of protein S-sulfenylation in cellsThe files in this archive represent the supporting data for "Site-specific mapping and quantification of protein S-sulfenylation in cells," a paper to appear in Nature Communications (2014). The authors are Jing Yang, Vinayak Gupta, Kate S. Carroll, and Daniel C. Liebler, working at Vanderbilt University School of Medicine in Nashville, TN, and The Scripps Research Institute, Jupiter, FL. Human cell lines were treated with an alkynyl probe to label S-sulfenylcysteines and the labeled proteins were tagged with an azidobiotin reagent by click chemistry, digested with trypsin, and desalted (described in the paper in detail). Peptide capture was accomplished through streptavidin columns. Data-dependent LC-MS/MS experiments without prior fractionation were conducted on a Thermo Q-Exactive, employing HCD fragmentation. Light and heavy variants of each peptide would carry 333.1689 Da and 339.2065 Da mass shifts on Cysteines, respectively, and iodoacetamide would modify remaining cysteines by 57.021464 Da. Thu Jul 10 13:02:00 BST 2014MSV00007877725175731