Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R22_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R23_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R26_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R27_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R29_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R21_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R28_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R29_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R25_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R24_03.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R26_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R24_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R25_01_B.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R22_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R23_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R21_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R21_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R25_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R28_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R26_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R27_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R27_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R28_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R24_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R22_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R23_02.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/20140514_CO_0347AnPe_R29_01.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2016/06/PXD004283/txt.zipprimaryOK200004990andreas.peschel@uni-tuebingen.deNicolas NalpasMass SpectrometryShotgun proteomicsNot availablePhenol-soluble modulinsExoproteomeQuantitative label-free proteomicsCulture supernatantLipopeptideStaphylococcus aureushttp://www.ebi.ac.uk/pride/archive/projects/PXD004283Three biological replicates of USA300 wild-type or mutant culture filtrates grown overnight in TSB were analysed. The same protein amount measured by Bradford assay in culture filtrates of the various strains was used for exoprotein precipitation with 10% ice-cold trichloroacetic acid overnight at 4°C. After centrifugation at 13,200 rpm at 4°C for 15 min the supernatants were discarded and precipitated proteins were air dried. For nanoLC-MS/MS analysis dried proteins were dissolved in a buffer containing 6 M urea, 2 M thiourea, 10 mM Tris at pH 8.0 and digested in solution. Peptide mixtures were then separated on an EasyLC nano-HPLC (Proxeon Biosystems) coupled to an LTQ Orbitrap Elite mass spectrometer (Thermo Fisher Scientific). Peptides were eluted with an 87-min segmented gradient of 5–33-90% HPLC solvent B (80% acetonitrile in 0.5% acetic acid). Each sample was run in technical triplicates.PrideNot availableNo PTMs are included in the datasetAcquired MS spectra were processed with the MaxQuant software package version 1.2.2.9 with integrated Andromeda search engine. Database searches were performed against a target decoy S. aureus USA300 database obtained from Uniprot, containing 5,295 protein entries and 248 commonly observed contaminants. The label-free algorithm was enabled as was the “match between runs” option. Label-free quantification (LFQ) protein intensities from the MaxQuant data output were used for relative protein quantification.ProteomicsAndreas PeschelLTQ Orbitrap ElitePARTIALInterfaculty Institute of Microbiology and Infection Medicine, Infection Biology Department, University of Tübingen, Auf der Morgenstelle 28 72076 Tübingen, GermanyStaphylococcus Aureus27470911 Hanzelmann D, Joo HS, Franz-Wachtel M, Hertlein T, Stevanovic S, Macek B, Wolz C, Götz F, Otto M, Kretschmer D, Peschel A. Toll-like receptor 2 activation depends on lipopeptide shedding by bacterial surfactants. Nat Commun. 2016 Jul 29;7:12304 10.1038/ncomms12304nalpas.nicolas@gmail.comBiologicalBiomedicalTuebingen UniversityGermanySepsis caused by Gram-positive bacterial pathogens is a major fatal disease but its molecular basis remains elusive. Toll-like receptor 2 (TLR2) has been implicated in the orchestration of inflammation and sepsis but its role appears to vary for different pathogen species and clones. Accordingly, Staphylococcus aureus clinical isolates differ substantially in their capacity to activate TLR2. Here we show that strong TLR2 stimulation depends on high-level production of phenol-soluble modulin (PSM) peptides in response to the global virulence activator Agr. PSMs are required for mobilizing lipoproteins, the TLR2 agonists, from the staphylococcal cytoplasmic membrane. Notably, the course of sepsis caused by PSM-deficient S. aureus is similar in wild-type and TLR2-deficient mice, but TLR2 is required for protection of mice against PSM-producing S. aureus. Thus, a crucial role of TLR2 depends on agonist release by bacterial surfactants. Modulation of this process may lead to new therapeutic strategies against Gram-positive infections.Toll-like receptor 2 activation depends on lipopeptide shedding by bacterial surfactants.Hanzelmann Dennis D, Joo Hwang-Soo HS, Franz-Wachtel Mirita M, Hertlein Tobias T, Stevanovic Stefan S, Macek Boris B, Wolz Christiane C, Götz Friedrich F, Otto Michael M, Kretschmer Dorothee D, Peschel Andreas AIL1BC, T22F8.160, MeCN, NCMe, determination, and rna binding 2, Vinegar, Chronic Airflow Obstructions, CASP-1, A4, CG7788, crice, Essigsaeure, chronic obstructive pulmonary disease (COPD), protein, Basodexan, Solvent, trichloro-, peptide, Polypeptides, Background, protein polypeptide chains, E260, ClvPrd, peptido, T22F8_160, Glacial Acetic Acid, ATGPR7, CH3-C#N, Cultural, chronic obstructive airways disease NOS (disorder), Chronic irreversible airway obstruction, Urea, GRP8, GRP7, Min, Chronic Obstructive Pulmonary Disease (COPD), protein aggregate, Il1bc, DmelCG42273, ethnicity, Divorced, COLD 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airflow limitation, cyanomethane, chronic obstructive airway disease, AU020952, Temperature, Separation, 10^[-9], DMDA1, E 260, uree, Dops, urea, Separations, CC1, GR-RBP7, GR-RBP8, IL-1 beta-converting enzyme, acide acetique, ethanenitrile, Interleukin-1 beta-converting enzyme, sample population, UREA, ME-IV, Interleukin-1 beta convertase, Protein Gene Products, Gene Proteins, MeCOOH, ACETIC ACID, SCARMD2, chronic obstructive lung disease (disorder), 3.4.22.36, Relativisms, cold (chronic obstructive lung disease), Peptid, INS No. 260, Chronic Obstructive Lung Disease, assay, acetic acid, Relativism, Cultural Relativism, Ethanoic acid, chronic obstructive pulmonary diseasesurfactants, CD282, Receptor, surfactant., TIL4, Toll/interleukin-1 receptor-like protein 4, activation, surface active agent, Toll Like Receptor 2, TLR2, TLR2 ReceptorApplications Software, Open Source, data, Computer Software, protein complex, T5E21.12, Search, protein, Computer, protein-containing complex, not 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Polypeptides, FOLH, peptido, diseases, Roles, responsivity, Concepts, Phenol, C530001K22Rik, diseases and disorders, Pyemia, 3.4.17.21, Staphlococcus pyogenes citreus, SH2-B, strong, Carbolic Acid, somitogenic mesoderm, viral infection, human disease, acide carbolique, inflammatory response, peptides, Circulating Lipoproteins, Membrane glutamate carboxypeptidase, Staphilococcus aureus, Karbolsaeure, Swiss Mice, Micrococcus aureus, virus process, NAALAD1, AUXIN TRANSPORT PROTEIN EIR1, Folylpoly-gamma-glutamate carboxypeptidase, inner endospore membrane, Phenylic acid, plasma membrane lipid bilayer, g, Carbol, plasmalemma, papilla, Role Concepts, WAVY ROOTS 6, Homo sapiens disease, anon-WO0140519.74, Phenylic alcohol, Phenic acid, ARABIDOPSIS THALIANA PIN-FORMED 2, anatomical protrusion, Blood Poisoning, unsegmented mesenchyme, mouse, lamina, flanges, Severe Sepsis, SH2-B PH domain-containing signaling mediator 1, acide phenique, PHENOL, cellular membrane, Role Concept, juxtamembrane, soluble, agonist, PSM, Psm, Toll Like Receptor 2, Mini-ICE, Diseases, shelf, Role, Pathogenicity, CG9538, bacterial inner membrane, presumptive somite mesoderm, Prostate-specific membrane antigen, Sepsis, Poisoning, TIL4, POLAR-AUXIN-TRANSPORT EFFLUX COMPONENT AGRAVITROPIC 1, TLR2, Phenolate, WAV6, Mus musculus, Pteroylpoly-gamma-glutamate carboxypeptidase, cell membrane, Caspase-14 subunit p10, Blood Poisonings, mice, FGCP, shelves, carbolic acid, Swiss Mouse, Infection and Infestation, Bloodstream Infections, Agr5, Caspase-14 subunit p19, MICE, Toll/interleukin-1 receptor-like protein 4, AGR1, projection, ridge, Bloodstream, surfactant, domesticus, Pro-rich, disease, Sodium Salt, DMDA, Staphylococcus pyogenes aureus, Poisonings, spine, Pyaemia, AGR, AGRAVITROPIC ROOT, Agr, surface active agent, PSMA, Lipoproteins, Mouse, Polypeptide, Innate, PIN-FORMED 2, 3.4.22.-, other disease, Inflammatory Response, SH2 domain-containing protein 1B, lamellae, agr, unsegmented paraxial mesoderm, surfactants, mini-ICE, Septicemias, somitomeric mesoderm, GCPII, process of organ, TYPE, protrusion, DAGA4, MUL3.3, lamella, Septicemia, resilient, House, tough, Sodium Phenolate, disease or disorder, N-acetylated-alpha-linked acidic dipeptidase I, Mus musculus domesticus, mKIAA1299, MUL3_3, MAM, phenol, SCG3, Mice, reactivity, SH2B, ETHYLENE INSENSITIVE ROOT 1, Benzenol, Swiss, Ly105, inflammation, mGCP, ridges, Infections and Infestations., RCB0750, non-neoplastic, Infestation and Infection, CD282, Innate Inflammatory Response, TLR2 Receptor, disorder, Oxybenzene, peptidos, species, Staphylococus aureus, PIN2, laminae, integral to plasma membrane, agonista, Lipoprotein, integral component of plasma membrane, anatomical process, disorders, Hydroxybenzene, Bloodstream Infection, medical condition, Peptide, GCP2, LGMD2C, Concept, ATPIN2, Glutamate carboxypeptidase II, polypeptide, Cell growth-inhibiting gene 27 protein, Mus, Infection, condition, agonists, Sodium, segmental plate, 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domesticus, mKIAA1299, MUL3_3, MAM, phenol, SCG3, Mice, reactivity, SH2B, ETHYLENE INSENSITIVE ROOT 1, Benzenol, Swiss, Ly105, inflammation, mGCP, ridges, Infections and Infestations., RCB0750, non-neoplastic, Infestation and Infection, CD282, Innate Inflammatory Response, TLR2 Receptor, disorder, Oxybenzene, peptidos, species, Staphylococus aureus, PIN2, laminae, integral to plasma membrane, agonista, Lipoprotein, integral component of plasma membrane, anatomical process, disorders, Hydroxybenzene, Bloodstream Infection, medical condition, Peptide, GCP2, LGMD2C, Concept, ATPIN2, Glutamate carboxypeptidase II, polypeptide, Cell growth-inhibiting gene 27 protein, Mus, Infection, condition, agonists, Sodium, segmental plate, Sh2bpsm1, flange, agoniste, organ process, AGRAVITROPIC ROOT 1, PH and SH2 domain-containing signaling mediator, Staphylococcus aureus subsp. anaerobius, DMDA1, PhOH, House Mice, Folate hydrolase 1, Streptococcus aureus, Laboratory Mice, processes, process, Micrococcus pyogenes, Carbolsaeure, Innate Inflammatory Responses, Phenolate Sodium, Pyohemias, AI425885, SCARMD2, processus, Peptid, virulence, Receptor, response, cytoplasmic membrane, Laboratory Mouse, Severesurfactants, CD282, Receptor, surfactant., TIL4, Toll/interleukin-1 receptor-like protein 4, activation, surface active agent, Toll Like Receptor 2, TLR2, TLR2 Receptor499trueToll-like receptor 2 activation depends on lipopeptide shedding by bacterial surfactantsSepsis caused by Gram-positive bacterial pathogens is a major fatal disease but its molecular basis remains elusive. Toll-like receptor 2 (TLR2) has been implicated in the orchestration of inflammation and sepsis but its role appears to vary for different pathogen species and clones. Accordingly, Staphylococcus aureus clinical isolates differ substantially in their capacity to activate TLR2. Here we show that strong TLR2 stimulation depends on high-level production of phenol-soluble modulin (PSM) peptides in response to the global virulence activator Agr. PSMs are required for mobilizing lipoproteins, the TLR2 agonists, from the staphylococcal cytoplasmic membrane. Notably, the course of sepsis caused by PSM-deficient S. aureus is similar in wild-type and TLR2-deficient mice, but TLR2 is required for protection of mice against PSM-producing S. aureus. Thus, a crucial role of TLR2 depends on agonist release by bacterial surfactants. Modulation of this process may lead to new therapeutic strategies against Gram-positive infections.2016-06-082016-06-06PXD00428344664499175582580388043330295358511145108061246196246197320637581136335090683635275838538791187947398007NCBITaxon:12805723071114301555714683514526236492364113546138858033888691568895475187043511232946928123977133103430436548135833925198183659219813143205914936329572637376938124781866132838828139545132949140816918045416193413903634530326442104117061274432108595614963674283166694369455972910039237514633087936944784208964317447127044597411946691454587932993529320101981264690410661145755121659939251216597216595160621213312831282128033952321832172920435315228112454349825747874980197525555362312951053948824261914548140479NCBITaxon:2157551534398920596876231935163098005888455818178081736309431796989361604885661409859566416242515059071187021187053204963063863235554320466322261866331299456586068845351216129567171360441001602358700264203694569410167634102107612773512793310029357861187002157641809699745774449156666754760445246763366411363367102398709NCBITaxon:40559235909278839652649821749821656915693498211872056971089448663661410NCBITaxon:49328715568910005894526808732311561263730391141554219328726872387248729872755529547624223141463055180066556484198822299767982510216998231174673897641155744689444474153NCBITaxon:50557960167767446859600653491182263129911457845863267656983556636983819201124136810893161001428099612NCBITaxon:548681546414307747946409860995865228319662776689605960615729513722187594467087562270861711013354834974138783430615470334249961574649987039202187827215407821190857075206189480418985350247158779627190252867463609646375939146247274612874601134506253452659197221963852638588596NCBITaxon:6157110517237263419610526419407722774694163481123869NCBITaxon:270361274767927459615325731326979699034694171344346360347515363296440421097606331492862879669968583558595968926071018568729411009118696141633316780781186941436183NCBITaxon:61911186988402317191186919694190802187878703847664553470122578656870861822607046183260705260707NCBITaxon:780NCBITaxon:1502451516658457702921158612184520928574707861921263854504457383379192875NCBITaxon:10239NCBITaxon:714282912680637044830064110555241590841228233446900815025754122810430085210977970671356225833136185509924NCBITaxon:5693826883064052779658334240906151015154067448242813076707087613837082816772850990119519533702492405360888175626153337602126719285321773119350184645177237111215323690147091571256274579031133355085962859631308212035332648373153NCBITaxon:9615318586141262200298493760833348333210184110065812375612666255749200360094153009411490352472517503983649908515151438992NCBITaxon:1155237453747445446415259311792866432359839062887051580119459932716016032932853422999810894535936457428135122511775058NCBITaxon:10407167971816738713491589103062915954126291585158493271595702100226128768913558893094514647920312421641332803828035192843439858089462179392118259028788929502790690174934272563227321890382928671247190400727537457114311233521140206411981087135547714245072295333555714154293157546224213921390114319338293550833292307411292211484896725264999757447275802405507366581214053576229412816078395763206672400540296543184318322465755499229575757592845933079727066714367331159556444214321381849223112109697622611117957529818184319233479866265281046604547479626782597477317513565050NCBITaxon:1313607699389424688795520148875197959621412890922853164351867631854316494089414424232NCBITaxon:1773214092115778929793244704207559101177499067499079913112273170187367451011610358132842626587210360983341392696113079813367953588931640710340153033373301581999253068839103372123368199404458611535711471612249113430531201755499351299385057317666964268845151345842157267872862386211151041607912983154689119835112552285625323213616763363168970413288818855116761080772326424861839376520965731116234273068808637721666898626NCBITaxon:2697049339862410312NCBITaxon:138997586448643319729176343863725673786391233435105231100905141405449317998686554055348654514776056810429863571246791884019373345087711982159739757429719427577985076255470256318623951655865886223864946454634076NCBITaxon:1107128835649184588NCBITaxon:592011459432676711116781077408440879778953496794408122690038039435128145953612351786162432303996686959785409640979541869420030895442432778697627912924995454090518042643040928692409320030228481231362542642815047510299388653862629034155405120897966287732396692026289955771371170089306218644195486575584628238235217362315577223994636010688420436010439947932184795444044731169681121279011622961731440772131693193128118361003610502318557984854318637499010041715916922596345910659210659033454288531844271716591037164784964102896883331249668425281094343197NCBITaxon:10357NCBITaxon:10114507601NCBITaxon:10359272634956029598802242752959526643732354432243081433619432749839642711913645169963112223360520121697927262327262412389938697286436803347471821314269704940324803029760381811477873401881274414153681639588858749402955468049224326408172128161405592042546382712744234342714081701274420127442649115437341316416601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