Identification of a collagen type I adhesin of Bacteroides fragilis

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dc.contributor.authorGalvão, Bruna P G Ven_ZA
dc.contributor.authorWeber, Brandon Wen_ZA
dc.contributor.authorRafudeen, Mohamed Sen_ZA
dc.contributor.authorFerreira, Eliane Oen_ZA
dc.contributor.authorPatrick, Sheilaen_ZA
dc.contributor.authorAbratt, Valerie Ren_ZA
dc.date.accessioned2015-11-10T14:48:45Z
dc.date.available2015-11-10T14:48:45Z
dc.date.issued2014en_ZA
dc.description.abstractBacteroides fragilis is an opportunistic pathogen which can cause life threatening infections in humans and animals. The ability to adhere to components of the extracellular matrix, including collagen, is related to bacterial host colonisation. Collagen Far Western analysis of the B. fragilis outer membrane protein (OMP) fraction revealed the presence two collagen adhesin bands of ∼31 and ∼34 kDa. The collagen adhesins in the OMP fraction were separated and isolated by two-dimensional SDS-PAGE and also purified by collagen affinity chromatography. The collagen binding proteins isolated by both these independent methods were subjected to tandem mass spectroscopy for peptide identification and matched to a single hypothetical protein encoded by B. fragilis NCTC 9343 (BF0586), conserved in YCH46 (BF0662) and 638R (BF0633) and which is designated in this study as cbp1 (collagen binding protein). Functionality of the protein was confirmed by targeted insertional mutagenesis of the cbp1 gene in B. fragilis GSH18 which resulted in the specific loss of both the ∼31 kDa and the ∼34 kDa adhesin bands. Purified his-tagged Cbp1, expressed in a B. fragilis wild-type and a glycosylation deficient mutant, confirmed that the cbp1 gene encoded the observed collagen adhesin, and showed that the 34 kDa band represents a glycosylated version of the ∼31 kDa protein. Glycosylation did not appear to be required for binding collagen. This study is the first to report the presence of collagen type I adhesin proteins in B. fragilis and to functionally identify a gene encoding a collagen binding protein.en_ZA
dc.identifier.apacitationGalvão, B. P. G. V., Weber, B. W., Rafudeen, M. S., Ferreira, E. O., Patrick, S., & Abratt, V. R. (2014). Identification of a collagen type I adhesin of Bacteroides fragilis. <i>PLoS One</i>, http://hdl.handle.net/11427/14838en_ZA
dc.identifier.chicagocitationGalvão, Bruna P G V, Brandon W Weber, Mohamed S Rafudeen, Eliane O Ferreira, Sheila Patrick, and Valerie R Abratt "Identification of a collagen type I adhesin of Bacteroides fragilis." <i>PLoS One</i> (2014) http://hdl.handle.net/11427/14838en_ZA
dc.identifier.citationGalvão, B. P., Weber, B. W., Rafudeen, M. S., Ferreira, E. O., Patrick, S., & Abratt, V. R. (2013). Identification of a collagen type I adhesin of Bacteroides fragilis. PloS one, 9(3), e91141. doi:10.1371/journal.pone.0091141en_ZA
dc.identifier.ris TY - Journal Article AU - Galvão, Bruna P G V AU - Weber, Brandon W AU - Rafudeen, Mohamed S AU - Ferreira, Eliane O AU - Patrick, Sheila AU - Abratt, Valerie R AB - Bacteroides fragilis is an opportunistic pathogen which can cause life threatening infections in humans and animals. The ability to adhere to components of the extracellular matrix, including collagen, is related to bacterial host colonisation. Collagen Far Western analysis of the B. fragilis outer membrane protein (OMP) fraction revealed the presence two collagen adhesin bands of ∼31 and ∼34 kDa. The collagen adhesins in the OMP fraction were separated and isolated by two-dimensional SDS-PAGE and also purified by collagen affinity chromatography. The collagen binding proteins isolated by both these independent methods were subjected to tandem mass spectroscopy for peptide identification and matched to a single hypothetical protein encoded by B. fragilis NCTC 9343 (BF0586), conserved in YCH46 (BF0662) and 638R (BF0633) and which is designated in this study as cbp1 (collagen binding protein). Functionality of the protein was confirmed by targeted insertional mutagenesis of the cbp1 gene in B. fragilis GSH18 which resulted in the specific loss of both the ∼31 kDa and the ∼34 kDa adhesin bands. Purified his-tagged Cbp1, expressed in a B. fragilis wild-type and a glycosylation deficient mutant, confirmed that the cbp1 gene encoded the observed collagen adhesin, and showed that the 34 kDa band represents a glycosylated version of the ∼31 kDa protein. Glycosylation did not appear to be required for binding collagen. This study is the first to report the presence of collagen type I adhesin proteins in B. fragilis and to functionally identify a gene encoding a collagen binding protein. DA - 2014 DB - OpenUCT DO - 10.1371/journal.pone.0091141 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2014 T1 - Identification of a collagen type I adhesin of Bacteroides fragilis TI - Identification of a collagen type I adhesin of Bacteroides fragilis UR - http://hdl.handle.net/11427/14838 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/14838
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0091141
dc.identifier.vancouvercitationGalvão BPGV, Weber BW, Rafudeen MS, Ferreira EO, Patrick S, Abratt VR. Identification of a collagen type I adhesin of Bacteroides fragilis. PLoS One. 2014; http://hdl.handle.net/11427/14838.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentDepartment of Molecular and Cell Biologyen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_ZA
dc.rights.holder© 2014 Galvão et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherCollagensen_ZA
dc.subject.otherAdhesinsen_ZA
dc.subject.otherOuter membrane proteinsen_ZA
dc.subject.otherSequence motif analysisen_ZA
dc.subject.otherMembrane proteinsen_ZA
dc.subject.otherGlycosylationen_ZA
dc.subject.otherPolymerase chain reactionen_ZA
dc.subject.otherPlasmid constructionen_ZA
dc.titleIdentification of a collagen type I adhesin of Bacteroides fragilisen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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