1887

Abstract

has been implicated in human periodontal diseases. It expresses a number of exoglycosidase enzymes capable of hydrolysing host proteoglycan residues. As a first stage to explore the role of these enzymes in periodontal tissue damage, the gene of W83, which encodes β--acetylhexosaminidase (β-Nahase), was cloned. The gene was expressed poorly in , but increased expression was achieved by cloning the gene downstream of the tac promoter. Southern blot analysis revealed that was present as a single copy, and it was found in all the other strains tested. In contrast, sequences homologous to were not detected in either or The gene was 2331 bp long and encoded a β-Nahase enzyme of 777 amino acids with a predicted molecular mass of 87 kDa. A characteristic signal peptide for an acylated lipoprotein was present at the amino-terminus, suggesting that the mature β-Nahase is a lipoprotein. The predicted amino acid sequence of the β-Nahase shared homology with the catalytic domains of the human β-Nahase enzyme and the chitinase of , suggesting a common catalytic mechanism.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-12-3399
1994-12-01
2021-10-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/12/mic-140-12-3399.html?itemId=/content/journal/micro/10.1099/13500872-140-12-3399&mimeType=html&fmt=ahah

References

  1. Bapat B., Ethier M., Noete K., Mahuran D., Gravel R.A. Cloning and sequence analysis of a cDNA encoding the ¡3-subunit of mouse zhexosaminidase. FEBS Lett 1987; 237:191–195
    [Google Scholar]
  2. Bartold P.M. Proteoglycans of the periodontum: structure, role and function. J Periodontal Res 1987; 22:431–444
    [Google Scholar]
  3. Bartold P.M., Weibkin O.W., Thornard J.C. Proteoglycans in human gingiva: molecular size distribution in epithelium and connective tissue. Arch Oral Biol 1982; 27:1–7
    [Google Scholar]
  4. Beighton D., Radford J.R., Taylor M.N. Glycosidase activities in gingival crevicular fluid in subjects with adult periodontitis and gingivitis. Arch Oral Biol 1992; 37:43–48
    [Google Scholar]
  5. Biggins M.D., Gibson T.J., Hong G.F. Buffer gradient gels and 35 S label as an aid to rapid DNA sequence determination. Proc Nail Acad Sci USA 1983; 80:3963–3965
    [Google Scholar]
  6. Birnboim H.C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 1979; 1:1513–1523
    [Google Scholar]
  7. Boureau H., Deere D., Carlier J.P., Guichet C., Bourlioux P. Identification of a Clostridium cocleatum strain involved in wcA-Clostridium difficile barrier effect and determination of its mucin degrading enzymes. Rer Microbiol 1993; 144:405–410
    [Google Scholar]
  8. Bourgeau G., Lapointe H., Peloquin P., Mayrand D. Cloning, expression and sequencing of a protease gene (tpr) from Porphyromonas gingivalis W83 in Escherichia coli. Infect Immun 1992; 60:3186–3192
    [Google Scholar]
  9. Brown C.A., Mahuran D.J. Active arginine residues in z-hexosaminidase. J Biol Chem 1991; 266:15855–15862
    [Google Scholar]
  10. Cabezas J.A. Some comments on the type references of official nomenclature (IUB) for β-N-acetylglucosaminidase, β-N-acetylhexosaminidase and β-IV-acetylgalactosaminidase. Biochem J 1989; 261:1059–1061
    [Google Scholar]
  11. Casaregola S., D'Ari R., Huisman O. Quantitative evaluation of recA gene expression in E. coli. Mol & Gen Genet 1982; 185:430–439
    [Google Scholar]
  12. Ciewell D.B., Helinski D.R. Supercoiled circular DNA-protein complex in Escherichia coli\ purification and conversion to an open circular form. Proc Natl Acad Sci USA 1969; 62:1159–1166
    [Google Scholar]
  13. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 1984; 12:387–395
    [Google Scholar]
  14. Dower W.G., Miller J.F., Ragsdale C.W. High efficiency transformation of Escherichia coli using electroporation. Nucleic Acids Res 1988; 16:6127–6145
    [Google Scholar]
  15. Feinberg A.P., Vogelstein B. A technique for radiolabelling restriction endonuclease fragments to a high specific activity. Anal Biochem 1983; 132:6–13
    [Google Scholar]
  16. Graham T.R., Zassenhaus P., Kaplan A. Molecular cloning of the cDNA which encodes the /LN-acetylhexosaminidase from Dictyostelium discoideum. J Biol Chem 1988; 263:16823–16829
    [Google Scholar]
  17. Greenman J., Minhas T. Production of cell-bound and vesicle-associated trypsin-like protease, alkaline phosphatase and Ar-acetyl-β-glucosaminidase by Bacteroides gingivalis W50. J Gen Microbiol 1989; 135:557–564
    [Google Scholar]
  18. Hayashi S., Wu H.C. Lipoproteins in bacteria. J Bioenerg Biomembr 1990; 22:451–471
    [Google Scholar]
  19. Higgins D.G., Bleasby A.J., Fuchs R. Clustal-V-improved software for multiple sequence alignment. Comput Appl Biosci 1992; 8:189–191
    [Google Scholar]
  20. Holt S.C., Bramanti T.E. Factors in virulence expression and their role in periodontal disease pathogenesis. Crit Rev Oral Biol Med 1991; 2:177–281
    [Google Scholar]
  21. Jensen H.K., Ledet T. Proteolysis of arterial basement membrane containing different amounts of carbohydrates. Thromb Res 1986; 44:47–53
    [Google Scholar]
  22. Kato T., Takahashi N., Kuramitsu H.K. Sequence analysis and characterisation of the Porphyromonas gingivalis prtC gene, which expresses a novel collagenase activity. J Bacteriol 1992; 174:3889–3895
    [Google Scholar]
  23. Koide N., Matso N., Muramatso T. Recognition of IgG by Fc receptor and complement: effects of glycosidase digestion. Biochem Biopbys Res Commun 1977; 75:838–844
    [Google Scholar]
  24. Korneluk R.G., Mahuran D.J., Neote K., Klavinis M.H., O'Dowd F., Tropak M., Willard H.F., Anderson M.-J., Lowden J.A., Gravel R.A. Isolation of cDNA clones coding for the a-subunit of human β-hexosaminidase. J Biol Chem 1986; 261:8407–8413
    [Google Scholar]
  25. Kytzia H.-J., Sandhoff K. Evidence for two different active sites on human z-hexosaminidase A. J Biol Chem 1985; 260:7568–7572
    [Google Scholar]
  26. Kytzia H.-J., Hinrichs U., Maire I., Suzuki K., Sandhoff K. Variant of GM2 gangliosidosis with hexosaminidase A having a severely changed substrate specificity. EMBO J 1983; 2:1201–1205
    [Google Scholar]
  27. Laughon B.E., f Syed S.A., Loesche W.J. API ZYM system for identification of Bacteroides spp., Capnocytophaga spp., and spirochetes of oral origin. J Clin Microbiol 1982; 15:97–102
    [Google Scholar]
  28. Li J., Ellen R.P., Hoover C.I., Felton J.R. Association of proteases of Porphyromonas (Bacteroides) gingivalis with its adhesion to Actinomyces viscosus. J Dent Res 1991; 70:82–86
    [Google Scholar]
  29. Lipman D.J., Pearson W.R. Rapid and sensitive protein similarity searches. Science 1985; 227:1435–1441
    [Google Scholar]
  30. Mayrand D., Holt S.C. Biology of asaccharolytic black-pigmented Bacteroides spp. Microbiol Rev 1988; 52:149–153
    [Google Scholar]
  31. McCarthy J.E.G., Gualerzi C. Translational control of prokaryotic gene expression. Trends Genet 1990; 6:78–85
    [Google Scholar]
  32. Milner J.S., Dymock D., Cooper R.M., Roberts I.S. Penicillin binding proteins from Erwinia amylovora: mutants lacking PBP2 are avirulent. J Bacteriol 1993; 175:6082–6088
    [Google Scholar]
  33. Nishikita M., Yoshimura F., Nodasaka Y. Possibility of Bacteroides gingivalis hemagglutinin possessing protease activity revealed by inhibition studies. Oral Microbiol Immunol 1989; 33:75–80
    [Google Scholar]
  34. O'Dowd B.F., Quan F., Willard H.F., Lamhonwah A.-M., Korneluk R.G., Lowden J.A., Gravel R.A., Mahuran D.J. Isolation of cDNA clones coding for the z-subunit of human β-hexosaminidase. Proc Natl Acad Sci USA 1985; 82:1184–1188
    [Google Scholar]
  35. Page R.C. The role of inflammatory mediators in the pathogenesis of periodontal disease. J Periodontal Res 1991; 26:230–242
    [Google Scholar]
  36. Park Y., & Mc Bride B. Characterisation of the tpr gene product and isolation of a specific protease-deficient mutant of Porphyromonas gingivalis W83. Infect Immun 1993; 61:4139–4146
    [Google Scholar]
  37. Pugsley A.P. The complete secretory pathway in Gramnegative bacteria. Microbiol Rev 1993; 57:50–108
    [Google Scholar]
  38. Rafael W.S., Zyskind J.W. The NjAT-diacetylchitobiase of Vibrio harveyi. J Biol Chem 1989; 264:14778–14783
    [Google Scholar]
  39. Roberts I.S., Mountford R., High N., Bitter-Suermann D., Jann K., Timmis K., Boulnois G.J. Molecular cloning and analysis of genes for the production of the K5, K7, K12, and K92 capsular polysaccharides in Escherichia coli. J Bacteriol 1986; 168:1228–1233
    [Google Scholar]
  40. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 1979; 13:319–354
    [Google Scholar]
  41. Rouslahti E. Proteoglycans in cell regulation. J Biol Chem 1989; 264:13369–13372
    [Google Scholar]
  42. Saito H., Miuria K.I. Preparation of transforming DNA by phenol treatment. Biochim Biophys Acta 1963; 72:619–629
    [Google Scholar]
  43. Sanger F., Nicklen S., Coulson A.R. DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 1977; 74:5463–5467
    [Google Scholar]
  44. Shah H.N., Collins M.D. Proposal for reclassification of Bacteroides asaccharolyticus, Bacteroides gingivalis and Bacteroides endo-dontalis in a new genus, Porphyromonas. Int J Syst Bacteriol 1988; 38:128–131
    [Google Scholar]
  45. Shah H.N., Gharbia S.H. Lysis of erythrocytes by the secreted cysteine proteinase of Porphyromonas gingivalis W83. FEMS Microbiol Lett 1989; 61:213–218
    [Google Scholar]
  46. Shah H.N., Williams R.A.D., Bowden G.H., Hardie J.M. Comparison of the biochemical properties of Bacteroides melaninogenicus from human dental plaque and other sites. J Appl Bacteriol 1976; 41:473–492
    [Google Scholar]
  47. Slots J., Genco R.I. Black-pigmented Bacteroides species, Capnocytophaga species and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonisation, survival and tissue destruction. J Dent Res 1984; 63:412–421
    [Google Scholar]
  48. Slots J., Bragd L., Wikstrom M., Dahlen G. The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal diseases in adults. J Clin Periodontal 1986; 13:570–577
    [Google Scholar]
  49. Smalley J.W., Birss A.J. Trypsi n-like enzyme activity of extracellular membrane vesicles of Bacteroides gingivalis W50. J Gen Microbiol 1987; 133:2883–2894
    [Google Scholar]
  50. Smith A.N., Boulnois G.J., Roberts I.S. Molecular analysis of the Escherichia coli K5 kps locus: identification and characterisation of an inner membrane capsular polysaccharide transport system. Mol Microbiol 1990; 4:1863–1869
    [Google Scholar]
  51. Stark M.J.R. Multicopy expression vectors carrying the lac repressor gene for regulated high level expression of genes in Escherichia coli. Gene 1987; 51:255–267
    [Google Scholar]
  52. Sundqvist G., Carlsson J., Herrmann B., Tarnvik A. Degradation of human immunoglobulins G and M and complement factors C3 and C5 by black-pigmented Bacteroides species. J Med Microbiol 1985; 19:85–94
    [Google Scholar]
  53. Takazoe I., Nakamura T., Okuda K. Colonisation of the subgingival region area by Bacteroides gingivalis. J Dent Ra 1984; 63:422–426
    [Google Scholar]
  54. Tipler L.S., Embery G. Glycosaminoglycan-depolymer-izing enzymes produced by anaerobic bacteria isolated from the human mouth. Arch Oral Biol 1985; 30:391–396
    [Google Scholar]
  55. Uitto V.-J. Extracellular matrix molecules and their receptors: an overview with special emphasis on periodontal tissues. Crit Rev Oral Biol Med 1991; 2:323–354
    [Google Scholar]
  56. Von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 1986; 14:4683–4690
    [Google Scholar]
  57. Watanabe T., Oyanagi W., Suzuki K., Ohnishi K., Tanaka H. Structure of the gene encoding chitinase D from Bacillus circulans WL-12 and possible homology of the enzyme to other prokaryotic chitinases and class III plant chitinases. I Bacteriol 1992; 174:408–414
    [Google Scholar]
  58. Werries E., Nebinger P., Franz A. Degradation of biogene oligosaccharides by yS-N-acetyl-hexosaminidase secreted by Entamoeba histolytica. Mol Biochem Parasitol 1983; 7:127–140
    [Google Scholar]
  59. White D., Mayrand D. Association of oral Bacteroides with gingivitis and adult periodontitis. J Periodontal Res 1981; 16:259–265
    [Google Scholar]
  60. Van Winkelhoff A.J., Van Steenbergen T.J.M., Kippuw N., De Graaf J. Further characterisation of Bacteroides endodontalis, an asaccharolytic black-pigmented Bacteroides species from oral cavity. J Clin Microbiol 1985; 22:75–79
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-12-3399
Loading
/content/journal/micro/10.1099/13500872-140-12-3399
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error