{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Msf":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD056157/PRF_F_2023_E_MART_1103_63780.msf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD056157/PRF_F_2023_E_MART_1103_63779.msf"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD056157/PRF_F_2023_E_MART_1103_63780.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD056157/PRF_F_2023_E_MART_1103_63779.raw"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["emartens@umich.edu"],"submitter":["Nicholas Pudlo"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":["Microbiota","Mucin","Bacteria"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD056157"],"sample_protocol":["After growing Hm in 25ml of DM-POG (n=2) to mid-log, cells were centrifuged, washed in PBS and frozen at -80°C. Supernatants were filtered through a 0.22µm filter and also stored at -80C. Both the cell pellet and supernatants were lyophilized prior to protein extraction.  Cell pellets were resuspended in 500µl lysis buffer (4% SDS, 50mM Tris, 10mM DTT, pH8.5), heated to 95°C for 10 min, cooled to room temperature and centrifuged for 2 min at 4,000 x g to removed cell debris/unlysed cells. Ice-cold acetone was added at four volumes to one volume of retained supernatants and precipitated overnight at -20°C. Proteins were centrifuged for 10 min at 4,000 x g and resulting pellets washed in 80% acetone prior to centrifuging at 17,000 x g for 10 min at 4°C. Supernatants were removed and the protein pellet allowed to air dry prior to resuspension in 75µl of proteomics buffer (8M Urea, 50mM HEPES, pH8). Approximately 10mls of freeze-dried supernatant solids were resuspended in 800µl of water and added to 800µl of PRMM solution (20% methanol, 0.05mM pyrogallol red, 0.16mM sodium molybdate, 1mM sodium oxalate, 50mM succinic acid, pH2) and adjusted to ~pH3 using HCl and pH test strips. Resulting solutions were incubated at RT for 2hrs and then 4°C overnight. Tubes were centrifuged at 10,000 x g for 60 min at 4°C and washed in 500µl ice-cold acetone. The centrifugation and acetone wash was repeated and air dried pellets resuspended in 30µl proteomics buffer (8M Urea, 50mM HEPES, pH8). Cell and supernatant protein dilutions were run on a 4-12% Bis-Tris NuPAGE  gel (Invitrogen) and subjected to silver staining (Invitrogen). Proteins were then submitted for Shotgun Proteomic analysis at the University of Michigan Proteomics Resource Facility using their optimized protocol.  Briefly, Cysteines were reduced by adding 50 l of 10 mM DTT and incubating at 45 C for 30 min.  Samples were cooled to room temperature and alkylation of cysteines was achieved by incubating with 65 mM 2-Chloroacetamide, under darkness, for 30 min at room temperature.  An overnight (~16 h) digestion with 1 ug sequencing grade, modified trypsin was carried out at 37 C with constant shaking in a Thermomixer.  Digestion was stopped by acidification and peptides were desalted using SepPak C18 cartridges using manufacturer’s protocol (Waters).  Samples were completely dried using vacufuge.  Resulting peptides were dissolved in 8 l of 0.1% formic acid/2% acetonitrile solution and 2 ls of the peptide solution were resolved on a nano-capillary reverse phase column (Acclaim PepMap C18, 2 micron, 50 cm, ThermoScientific) using a 0.1% formic acid/2% acetonitrile (Buffer A) and 0.1% formic acid/95% acetonitrile (Buffer B) gradient at 300 nl/min over a period of 90 min.  Eluent was directly introduced into Orbitrap Fusion tribrid mass spectrometer (Thermo Scientific, San Jose CA) using an EasySpray source.  MS1 scans were acquired at 120K resolution (AGC target=2x105; max IT=100 ms).  Data-dependent High-energy C-trap dissociation MS/MS spectra were acquired using Top speed method (3 seconds) following each MS1 scan (NCE ~32%; AGC target 5x104; max IT 50 ms, 15K resolution)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Proteins were identified by searching the MS/MS data against UGRO018 database (3464 entries) using Proteome Discoverer (v3, Thermo Scientific).  Search parameters included MS1 mass tolerance of 10 ppm and fragment tolerance of 0.2 Da; two missed cleavages were allowed; carbamidimethylation of cysteine was considered fixed modification and oxidation of methionine, deamidation of aspergine and glutamine were considered as potential modifications.  False discovery rate (FDR) was determined using Percolator and proteins/peptides with a FDR of ≤1% were retained for further analysis."],"omics_type":["Proteomics"],"labhead":["Eric C Martens"],"instrument_platform":[""],"labhead_affiliation":["Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI USA."],"submission_type":["PARTIAL"],"species":["Erysipelotrichaceae Bacterium"],"publication":["41856288 Pudlo NA, Pereira GV, Schaus SR, Yang Q, Fuentes JJ, Jin C, Hein R, Zhang L, Terrapon N, Lyssiotis CA, Schmidt TM, Hansson GC, Luis AS, Martens EC. The butyrate-producing Gram-positive human gut bacterium, Hoskinsella mucinilytica, selectively targets host mucin N-acetylhexosamines. J Biol Chem. 2026 302(5):111371 10.1016/j.jbc.2026.111371"],"submitter_mail":["npudlo@umich.edu"],"submitter_affiliation":["University of Michigan"],"submitter_country":["United States"],"pubmed_abstract":["The personalized microbial communities (micobiota) that inhabit the distal guts of humans have evolved to process a variety of complex carbohydrates. Many gut bacteria depolymerize and ferment dietary fiber polysaccharides, mutualistically providing short-chain fatty acids and other metabolites to their host. Some human gut bacteria have evolved to utilize components of host mucin glycoproteins-the major component of secreted mucus that protects the gut. Recent studies have implicated certain mucin-degrading bacteria in the development of intestinal inflammation, making the identification of new gut bacteria that possess the ability to degrade mucins an important goal. We used gastric mucin to isolate a novel bacterium, Hoskinsella mucinilytica, that is largely restricted to using the GlcNAc and GalNAc sugars found in the O-linked glycans appended to secreted mucin. This butyrate-producing bacterium accesses these sugars from both polymeric gastric mucin and chemically released oligosaccharides. It also has a genome with correspondingly restricted carbohydrate-active enzyme content, with only three demonstrated mucin-degrading glycoside hydrolase enzymes belonging to families GH31, 36, and 89. Surprisingly, strains with an identical 16S rRNA V4 region to this isolate appear to be rare in the now numerous sequence-based microbiota surveys, with only 30 of 7390 (0.41%) human subjects harboring detectable levels of this bacterium in their stool, with an overall relative abundance ranging from 0.0004% to 0.013% when it is detected. This combination of low prevalence and low abundance suggests that this species could occupy an unknown niche for which access to mucin is important but otherwise renders it difficult to detect in stool-based microbiota surveys."],"pubmed_title":["The butyrate-producing Gram-positive human gut bacterium, Hoskinsella mucinilytica, selectively targets host mucin N-acetylhexosamines."],"pubmed_authors":["Pudlo Nicholas A NA, Pereira Gabriel Vasconcelos GV, Schaus Sadie R SR, Yang Qinnan Q, Fuentes Jaime J JJ, Jin Chunsheng C, Hein Robert R, Zhang Li L, Terrapon Nicolas N, Lyssiotis Costas A CA, Schmidt Thomas M TM, Hansson Gunnar C GC, Luis Ana S AS, Martens Eric C EC"],"additional_accession":[]},"is_claimable":false,"name":"The butyrate-producing Gram-positive human gut bacterium, Hoskinsella mucinilytica, selectively targets host mucin N-acetylhexosamines","description":"The personalized communities of microorganisms (microbiota) that inhabit the distal guts of humans have evolved to process a variety of complex carbohydrates. Many gut bacteria depolymerize and ferment dietary fiber polysaccharides, mutualistically providing short-chain fatty-acids to their host. Some human gut bacteria have evolved to utilize components of host mucin glycoproteins—the major component of protective mucus—and recent studies have implicated specific mucin-degrading commensal bacteria in the development of intestinal inflammation. Thus, identifying new gut bacteria that possess the ability to degrade host mucins and determining how these organisms might cooperate or compete for mucin components during health and disease is an important goal. Here, we used culture on solid medium containing gastric mucin as the main carbon source to isolate a novel bacterium that is largely restricted to using the N-acetylglucosamine and N-acetylgalactosamine sugars found in mucin O-glycans. This butyrate-producing bacterium accesses these sugars from both polymeric gastric mucin and chemically released oligosaccharides and has a genome with correspondingly restricted carbohydrate-active enzyme content. Surprisingly, strains related to this isolate appear to be rare in the now numerous sequence-based microbiota surveys with only 32/3781 (0.85%) human subjects containing this bacterium with an overall relative abundance ranging from 0.004-0.057%. This combination of low prevalence and abundance, suggests that this species could occupy an unknown niche in which access to mucin is important but otherwise render it difficult to detect in mostly feces-based microbiota surveys.","dates":{"publication":"2026-04-27","submission":"2024-09-24"},"accession":"PXD056157","cross_references":{"TAXONOMY":["NEWT:108018","NEWT:377960","NEWT:1131","NEWT:190486","NEWT:2737160","NEWT:2042546","NEWT:148942","NEWT:295546","NEWT:258594","NEWT:1126","NEWT:1129","NEWT:89187","NEWT:309807","NEWT:89184","NEWT:309800","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:44270","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:93217","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:10034","NEWT:661410","NEWT:488003","NEWT:224326","NEWT:376619","NEWT:3765","NEWT:2433","NEWT:3767","NEWT:433727","NEWT:135622","NEWT:672127","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:3760","NEWT:36630","NEWT:381198","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:44250","NEWT:317447","NEWT:698761","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:11308","NEWT:12637","NEWT:2164133","NEWT:1180","NEWT:2767846","NEWT:1343074","NEWT:314344","NEWT:108061","NEWT:224308","NEWT:3134131","NEWT:49928","NEWT:160621","NEWT:110368","NEWT:1916073","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:1310165","NEWT:114727","NEWT:5516","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:1410384","NEWT:1310173","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:135666","NEWT:419481","NEWT:5507","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:709964","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:65121","NEWT:3716","NEWT:3715","NEWT:2065263","NEWT:1549675","NEWT:177437","NEWT:9014","NEWT:10891","NEWT:118221","NEWT:34869","NEWT:226126","NEWT:1616117","NEWT:34865","NEWT:550537","NEWT:284812","NEWT:284811","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:1246791","NEWT:34875","NEWT:1431297","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:1883426","NEWT:657321","NEWT:1423687","NEWT:657322","NEWT:10886","NEWT:3708","NEWT:536231","NEWT:219813","NEWT:1301262","NEWT:319224","NEWT:2305479","NEWT:215451","NEWT:460519","NEWT:272843","NEWT:272844","NEWT:1348799","NEWT:55784","NEWT:485","NEWT:2499525","NEWT:467144","NEWT:142786","NEWT:28196","NEWT:2853422","NEWT:487","NEWT:1679718","NEWT:387344","NEWT:657318","NEWT:480","NEWT:67767","NEWT:46835","NEWT:216778","NEWT:376686","NEWT:237610","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:141469","NEWT:272831","NEWT:987059","NEWT:491","NEWT:422203","NEWT:460523","NEWT:2423","NEWT:85694","NEWT:85692","NEWT:911045","NEWT:3750","NEWT:85698","NEWT:291146","NEWT:233271","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:415978","NEWT:410289","NEWT:191701","NEWT:9054","NEWT:472","NEWT:29459","NEWT:34839","NEWT:476","NEWT:28128","NEWT:470","NEWT:41122","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:63363","NEWT:3735","NEWT:29491","NEWT:911022","NEWT:446","NEWT:153481","NEWT:714485","NEWT:445","NEWT:2003758","NEWT:29486","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:3726","NEWT:30455","NEWT:28141","NEWT:9030","NEWT:9031","NEWT:3728","NEWT:108931","NEWT:219799","NEWT:1325292","NEWT:219798","NEWT:548","NEWT:150475","NEWT:546","NEWT:1431340","NEWT:256737","NEWT:51350","NEWT:8694","NEWT:8692","NEWT:542","NEWT:394503","NEWT:13076","NEWT:219785","NEWT:451515","NEWT:8673","NEWT:550","NEWT:10804","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:40686","NEWT:1225786","NEWT:771870","NEWT:8665","NEWT:51338","NEWT:377142","NEWT:9995","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:10832","NEWT:311339","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:2585209","NEWT:643680","NEWT:214092","NEWT:40697","NEWT:13095","NEWT:162425","NEWT:242124","NEWT:9986","NEWT:8654","NEWT:536","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:80833","NEWT:479096","NEWT:28108","NEWT:28104","NEWT:1114957","NEWT:568703","NEWT:568708","NEWT:51315","NEWT:52641","NEWT:353152","NEWT:116597","NEWT:353153","NEWT:116598","NEWT:227377","NEWT:1774284","NEWT:27689","NEWT:207769","NEWT:40674","NEWT:51329","NCBITaxon:1763","NEWT:443906","NEWT:13037","NEWT:519","NEWT:941638","NEWT:511","NEWT:7394","NEWT:6063","NEWT:404330","NEWT:1328388","NEWT:13064","NEWT:529120","NEWT:300641","NEWT:39655","NEWT:206411","NEWT:80869","NEWT:214053","NEWT:80863","NEWT:665890","NEWT:52639","NEWT:52638","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:1196866","NEWT:6039","NEWT:37003","NEWT:1045775","NEWT:1123869","NCBITaxon:1335626","NEWT:7370","NEWT:280406","NEWT:83906","NEWT:99488","NEWT:99487","NEWT:1134506","NEWT:255470","NEWT:762903","NEWT:27697","NEWT:86185","NEWT:9935","NEWT:266940","NEWT:287889","NEWT:9938","NEWT:1028307","NEWT:52603","NEWT:92867","NEWT:92866","NEWT:1344414","NEWT:202950","NEWT:741158","NEWT:1293091","NEWT:202956","NEWT:483179","NEWT:315750","NEWT:99490","NEWT:632957","NEWT:265619","NEWT:11294","NEWT:754466","NEWT:9925","NC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