title: The modular xylanase Xyn10A from Rhodothermus marinus is cell-attached, and its C-terminal domain has several putative homologues among cell-attached proteins within the phylum Bacteroidetes.
abstract: Until recently, the function of the fifth domain of the thermostable modular xylanase Xyn10A from Rhodothermus marinus was unresolved. A putative homologue to this domain was however identified in a mannanase (Man26A) from the same microorganism which raised questions regarding a common function. An extensive search of all accessible data-bases as well as the partially sequenced genomes of R. marinus and Cytophaga hutchinsonii showed that homologues of this domain were encoded by multiple genes in microorganisms in the phylum Bacteroidetes. Moreover, the domain occurred invariably at the C-termini of proteins that were predominantly extra-cellular/cell attached. A primary structure motif of three conserved regions including structurally important glycines and a proline was also identified suggesting a conserved 3D fold. This bioinformatic evidence suggested a possible role of this domain in mediating cell attachment. To confirm this theory, R. marinus was grown, and activity assays showed that the major part of the xylanase activity was connected to whole cells. Moreover, immunocytochemical detection using a Xyn10A-specific antibody proved presence of Xyn10A on the R. marinus cell surface. In the light of this, a revision of experimental data present on both Xyn10A and Man26A was performed, and the results all indicate a cell-anchoring role of the domain, suggesting that this domain represents a novel type of module that mediates cell attachment in proteins originating from members of the phylum Bacteroidetes.
authors.affiliation: Department Biotechnology, Center for Chemistry and Chemical engineering, Lund University, P.O.Box 124, SE-221 00 Lund, Sweden. eva.nordberg_karlsson@biotek.lu.se
authors.forename: Eva Nordberg
authors.initials: EN
authors.lastname: Karlsson
authors.forename: Maher Abou
authors.initials: MA
authors.lastname: Hachem
authors.forename: Santosh
authors.initials: S
authors.lastname: Ramchuran
authors.forename: Hugo
authors.initials: H
authors.lastname: Costa
authors.forename: Olle
authors.initials: O
authors.lastname: Holst
authors.forename: Åsa
authors.initials: Å
authors.lastname: Fex Svenningsen
authors.forename: Gudmundur O
authors.initials: GO
authors.lastname: Hreggvidsson
mesh_terms: D000595:Amino Acid Sequence; D041963:Bacteroidetes / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D002473:Cell Wall / Q000201:enzymology*; D019295:Computational Biology / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010802:Phylogeny; D045245:Rhodothermus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D043325:Xylan Endo-1,3-beta-Xylosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D043325:Xylan Endo-1,3-beta-Xylosidase