title: Streptococcus mutans SMU.623c codes for a functional, metal-dependent polysaccharide deacetylase that modulates interactions with salivary agglutinin.
abstract: The genome sequence of the oral pathogen Streptococcus mutans predicts the presence of two putative polysaccharide deacetylases. The first, designated PgdA in this paper, shows homology to the catalytic domains of peptidoglycan deacetylases from Streptococcus pneumoniae and Listeria monocytogenes, which are both thought to be involved in the bacterial defense mechanism against human mucosal lysozyme and are part of the CAZY family 4 carbohydrate esterases. S. mutans cells in which the pgdA gene was deleted displayed a different colony texture and a slightly increased cell surface hydrophobicity and yet did not become hypersensitive to lysozyme as shown previously for S. pneumoniae. To understand this apparent lack of activity, the high-resolution X-ray structure of S. mutans PgdA was determined; it showed the typical carbohydrate esterase 4 fold, with metal bound in a His-His-Asp triad. Analysis of the protein surface showed that an extended groove lined with aromatic residues is orientated toward the active-site residues. The protein exhibited metal-dependent de-N-acetylase activity toward a hexamer of N-acetylglucosamine. No activity was observed toward shorter chitooligosaccharides or a synthetic peptidoglycan tetrasaccharide. In agreement with the lysozyme data this would suggest that S. mutans PgdA does not act on peptidoglycan but on an as-yet-unidentified polysaccharide within the bacterial cell surface. Strikingly, the pgdA-knockout strain showed a significant increase in aggregation/agglutination by salivary agglutinin, in agreement with this gene acting as a deacetylase of a cell surface glycan.
authors.affiliation: Department of Cariology Endodontology Pedodontology, ACTA, Louwesweg 1, 1066 EA Amsterdam, The Netherlands. d.deng@acta.nl
authors.forename: Dong Mei
authors.initials: DM
authors.lastname: Deng
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Urch
authors.forename: Jacob M
authors.initials: JM
authors.lastname: ten Cate
authors.forename: Vincenzo A
authors.initials: VA
authors.lastname: Rao
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
authors.forename: Wim
authors.initials: W
authors.lastname: Crielaard
mesh_terms: D000373:Agglutinins / Q000378:metabolism* / Q000502:physiology; D000581:Amidohydrolases / Q000172:deficiency / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D002686:Chitin / Q000502:physiology; D002876:Chromosomes, Bacterial / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D055786:Gene Knockout Techniques; D020022:Genetic Predisposition to Disease; D008670:Metals / Q000494:pharmacology; D009113:Muramidase / Q000378:metabolism; D009844:Oligosaccharides / Q000737:chemistry; D010457:Peptidoglycan / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012463:Saliva / Q000382:microbiology* / Q000502:physiology; D013295:Streptococcus mutans / Q000201:enzymology* / Q000235:genetics*
chemical_list: D000373:Agglutinins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008670:Metals; D009844:Oligosaccharides; D010457:Peptidoglycan; D002686:Chitin; D009113:Muramidase; D000581:Amidohydrolases; C022427:polysaccharide deacetylase