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dc.contributor.authorWei, Guoxianen_US
dc.contributor.authorTian, Naen_US
dc.contributor.authorValery, Adriana C.en_US
dc.contributor.authorZhong, Yien_US
dc.contributor.authorSchuppan, Detlefen_US
dc.contributor.authorHelmerhorst, Eva Jen_US
dc.coverage.spatialUnited Statesen_US
dc.date2015-02-01
dc.date.accessioned2020-05-19T15:05:18Z
dc.date.available2020-05-19T15:05:18Z
dc.date.issued2015-06
dc.identifierhttps://www.ncbi.nlm.nih.gov/pubmed/25895519
dc.identifier.citationGuoxian Wei, Na Tian, Adriana C Valery, Yi Zhong, Detlef Schuppan, Eva J Helmerhorst. 2015. "Identification of pseudolysin (lasB) as an aciduric gluten-degrading enzyme with high therapeutic potential for celiac disease." Am J Gastroenterol, Volume 110, Issue 6, pp. 899 - 908. https://doi.org/10.1038/ajg.2015.97
dc.identifier.issn1572-0241
dc.identifier.urihttps://hdl.handle.net/2144/41016
dc.descriptionPublished in final edited form as: Am J Gastroenterol. 2015 June; 110(6): 899–908. doi:10.1038/ajg.2015.97.en_US
dc.description.abstractOBJECTIVES Immunogenic gluten proteins implicated in celiac disease (CD) largely resist degradation by human digestive enzymes. Here we pursued the isolation of gluten-degrading organisms from human feces, aiming at bacteria that would digest gluten under acidic conditions, as prevails in the stomach. METHODS Bacteria with gluten-degrading activities were isolated using selective gluten agar plates at pH 4.0 and 7.0. Proteins in concentrated bacterial cell sonicates were separated by diethylaminoethanol chromatography. Enzyme activity was monitored with chromogenic substrates and gliadin zymography. Elimination of major immunogenic gluten epitopes was studied with R5 and G12 enzyme-linked immunosorbent assays. RESULTS Gliadin-degrading enzyme activities were observed for 43 fecal isolates, displaying activities in the ~150-200 and <50 kDa regions. The active strains were identified as Pseudomonas aeruginosa. Gliadin degradation in gel was observed from pH 2.0 to 7.0. Liquid chromatography-electrospray ionization-tandem mass spectrometry analysis identified the enzyme as pseudolysin (lasB), a metalloprotease belonging to the thermolysin (M4) family proteases. Its electrophoretic mobility in SDS-polyacrylamide gel electrophoresis and gliadin zymogram gels was similar to that of a commercial lasB preparation, with tendency of oligomerization. Pseudolysin eliminated epitopes recognized by the R5 antibody, while those detected by the G12 antibody remained intact, despite destruction of the nearby major T-cell epitope QPQLPY. CONCLUSIONS Pseudolysin was identified as an enzyme cleaving gluten effectively at extremely low as well as near-neutral pH values. The potential to degrade gluten during gastric transport opens possibilities for its application as a novel therapeutic agent for the treatment of CD.en_US
dc.description.sponsorshipK02 AI101067 - NIAID NIH HHS; R01 AI087803 - NIAID NIH HHS; AI087803 - NIAID NIH HHS; AI101067 - NIAID NIH HHSen_US
dc.description.urihttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=25895519
dc.format.extentp. 899 - 908en_US
dc.languageeng
dc.language.isoen_US
dc.publisherWolters Kluwer Health, Inc.en_US
dc.relation.ispartofThe American Journal of Gastroentorology
dc.rightsAttribution 4.0 Internationalen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBacterial proteinsen_US
dc.subjectCeliac diseaseen_US
dc.subjectChromatography, high pressure liquiden_US
dc.subjectChromatography, ion exchangeen_US
dc.subjectElectrophoresis, polyacrylamide gelen_US
dc.subjectEnzyme-linked immunosorbent assayen_US
dc.subjectFecesen_US
dc.subjectGliadinen_US
dc.subjectGlutensen_US
dc.subjectHumansen_US
dc.subjectHydrogen-ion concentrationen_US
dc.subjectMetalloendopeptidasesen_US
dc.subjectMicrobiotaen_US
dc.subjectSpectrometry, mass, electrospray ionizationen_US
dc.subjectTandem mass spectrometryen_US
dc.subjectClinical sciencesen_US
dc.subjectGastroenterology & hepatologyen_US
dc.titleIdentification of pseudolysin (lasB) as an aciduric gluten-degrading enzyme with high therapeutic potential for celiac diseaseen_US
dc.typeArticleen_US
dc.description.versionAccepted manuscripten_US
dc.identifier.doi10.1038/ajg.2015.97
pubs.elements-sourcepubmeden_US
pubs.notesEmbargo: Not knownen_US
pubs.organisational-groupBoston Universityen_US
pubs.organisational-groupBoston University, College of Health & Rehabilitation Sciences: Sargent Collegeen_US
pubs.organisational-groupBoston University, College of Health & Rehabilitation Sciences: Sargent College, Physical Therapy and Athletic Trainingen_US
pubs.organisational-groupBoston University, Henry M. Goldman School of Dental Medicineen_US
pubs.publication-statusPublisheden_US
dc.identifier.mycv40997


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Except where otherwise noted, this item's license is described as Attribution 4.0 International