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. 1997 Jun;65(6):2292–2298. doi: 10.1128/iai.65.6.2292-2298.1997

Effects of antibodies against cell surface protein antigen PAc-glucosyltransferase fusion proteins on glucan synthesis and cell adhesion of Streptococcus mutans.

H Yu 1, Y Nakano 1, Y Yamashita 1, T Oho 1, T Koga 1
PMCID: PMC175318  PMID: 9169766

Abstract

Cell surface protein antigen (PAc) and glucosyltransferases (GTFs) produced by Streptococcus mutans are considered to be major colonization factors of the organism, and the inhibition of these two factors is predicted to provide protection against dental caries. In this study, we have constructed fusion protein PAcA-GB, a fusion of the saliva-binding alanine-rich region (PAcA) of PAc with the glucan binding (GB) domain of GTF-I, an enzyme catalyzing the synthesis of water-insoluble glucan from sucrose, and fusion protein PAcA-SB, a fusion of PAcA with the sucrose binding (SB) domain of GTF-I. The recombinant fusion proteins were purified from cell extracts of Escherichia coli harboring the fusion genes, and rabbit antibodies against these fusion proteins were prepared. Water-insoluble glucan synthesis by cell-associated and cell-free GTF preparations from S. mutans as well as total glucan synthesis by GTF-I was markedly inhibited by anti-PAcA-GB immunoglobulin G (IgG) antibodies but not by anti-PAcA-SB IgG antibodies. Significant inhibition of the sucrose-independent and sucrose-dependent adhesion of S. mutans to saliva-coated hydroxyapatite beads was observed when anti-PAcA-GB antibodies were added to the reaction mixture. Anti-PAcA-SB antibodies inhibited the adhesion of S. mutans to the beads in the absence of sucrose but not in the presence of sucrose. Immunization with the fusion protein PAcA-GB may be useful for controlling the colonization of teeth by S. mutans.

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Selected References

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  1. Amann E., Ochs B., Abel K. J. Tightly regulated tac promoter vectors useful for the expression of unfused and fused proteins in Escherichia coli. Gene. 1988 Sep 30;69(2):301–315. doi: 10.1016/0378-1119(88)90440-4. [DOI] [PubMed] [Google Scholar]
  2. Aoki H., Shiroza T., Hayakawa M., Sato S., Kuramitsu H. K. Cloning of a Streptococcus mutans glucosyltransferase gene coding for insoluble glucan synthesis. Infect Immun. 1986 Sep;53(3):587–594. doi: 10.1128/iai.53.3.587-594.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowen W. H., Schilling K., Giertsen E., Pearson S., Lee S. F., Bleiweis A., Beeman D. Role of a cell surface-associated protein in adherence and dental caries. Infect Immun. 1991 Dec;59(12):4606–4609. doi: 10.1128/iai.59.12.4606-4609.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brady L. J., Piacentini D. A., Crowley P. J., Oyston P. C., Bleiweis A. S. Differentiation of salivary agglutinin-mediated adherence and aggregation of mutans streptococci by use of monoclonal antibodies against the major surface adhesin P1. Infect Immun. 1992 Mar;60(3):1008–1017. doi: 10.1128/iai.60.3.1008-1017.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chia J. S., Lin R. H., Lin S. W., Chen J. Y., Yang C. S. Inhibition of glucosyltransferase activities of Streptococcus mutans by a monoclonal antibody to a subsequence peptide. Infect Immun. 1993 Nov;61(11):4689–4695. doi: 10.1128/iai.61.11.4689-4695.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cope P. A., Mooser G. Antibodies against active-site peptides common to glucosyltransferases of mutans streptococci. Infect Immun. 1993 Nov;61(11):4814–4817. doi: 10.1128/iai.61.11.4814-4817.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crowley P. J., Brady L. J., Piacentini D. A., Bleiweis A. S. Identification of a salivary agglutinin-binding domain within cell surface adhesin P1 of Streptococcus mutans. Infect Immun. 1993 Apr;61(4):1547–1552. doi: 10.1128/iai.61.4.1547-1552.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dertzbaugh M. T., Elson C. O. Comparative effectiveness of the cholera toxin B subunit and alkaline phosphatase as carriers for oral vaccines. Infect Immun. 1993 Jan;61(1):48–55. doi: 10.1128/iai.61.1.48-55.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gangloff S., M'Zoughi R., Lett E., Scholler M., Baer J., Pini A., Ogier J., Klein J. P. Epitope mapping of Streptococcus mutans SR protein and human IgG cross-reactive determinants, by using recombinant proteins and synthetic peptides. J Immunol. 1992 May 15;148(10):3249–3255. [PubMed] [Google Scholar]
  10. Giffard P. M., Jacques N. A. Definition of a fundamental repeating unit in streptococcal glucosyltransferase glucan-binding regions and related sequences. J Dent Res. 1994 Jun;73(6):1133–1141. doi: 10.1177/00220345940730060201. [DOI] [PubMed] [Google Scholar]
  11. Hajishengallis G., Hollingshead S. K., Koga T., Russell M. W. Mucosal immunization with a bacterial protein antigen genetically coupled to cholera toxin A2/B subunits. J Immunol. 1995 May 1;154(9):4322–4332. [PubMed] [Google Scholar]
  12. Hajishengallis G., Koga T., Russell M. W. Affinity and specificity of the interactions between Streptococcus mutans antigen I/II and salivary components. J Dent Res. 1994 Sep;73(9):1493–1502. doi: 10.1177/00220345940730090301. [DOI] [PubMed] [Google Scholar]
  13. Hamada S., Horikoshi T., Minami T., Okahashi N., Koga T. Purification and characterization of cell-associated glucosyltransferase synthesizing water-insoluble glucan from serotype c Streptococcus mutans. J Gen Microbiol. 1989 Feb;135(Pt 2):335–344. doi: 10.1099/00221287-135-2-335. [DOI] [PubMed] [Google Scholar]
  14. Hanada N., Kuramitsu H. K. Isolation and characterization of the Streptococcus mutans gtfD gene, coding for primer-dependent soluble glucan synthesis. Infect Immun. 1989 Jul;57(7):2079–2085. doi: 10.1128/iai.57.7.2079-2085.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kelly C. G., Todryk S., Kendal H. L., Munro G. H., Lehner T. T-cell, adhesion, and B-cell epitopes of the cell surface Streptococcus mutans protein antigen I/II. Infect Immun. 1995 Sep;63(9):3649–3658. doi: 10.1128/iai.63.9.3649-3658.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kelly C., Evans P., Bergmeier L., Lee S. F., Progulske-Fox A., Harris A. C., Aitken A., Bleiweis A. S., Lehner T. Sequence analysis of the cloned streptococcal cell surface antigen I/II. FEBS Lett. 1989 Nov 20;258(1):127–132. doi: 10.1016/0014-5793(89)81632-1. [DOI] [PubMed] [Google Scholar]
  17. Koga T., Asakawa H., Okahashi N., Hamada S. Sucrose-dependent cell adherence and cariogenicity of serotype c Streptococcus mutans. J Gen Microbiol. 1986 Oct;132(10):2873–2883. doi: 10.1099/00221287-132-10-2873. [DOI] [PubMed] [Google Scholar]
  18. Koga T., Okahashi N., Takahashi I., Kanamoto T., Asakawa H., Iwaki M. Surface hydrophobicity, adherence, and aggregation of cell surface protein antigen mutants of Streptococcus mutans serotype c. Infect Immun. 1990 Feb;58(2):289–296. doi: 10.1128/iai.58.2.289-296.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Koga T., Yamashita Y., Nakano Y., Kawasaki M., Oho T., Yu H., Nakai M., Okahashi N. Surface proteins of Streptococcus mutans. Dev Biol Stand. 1995;85:363–369. [PubMed] [Google Scholar]
  20. Kuramitsu H. K., Smorawinska M., Nakano Y. J., Shimamura A., Lis M. Analysis of glucan synthesis by Streptococcus mutans. Dev Biol Stand. 1995;85:303–307. [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Laloi P., Munro C. L., Jones K. R., Macrina F. L. Immunologic characteristics of a Streptococcus mutans glucosyltransferase B sucrose-binding site peptide-cholera toxin B-subunit chimeric protein. Infect Immun. 1996 Jan;64(1):28–36. doi: 10.1128/iai.64.1.28-36.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee S. F., Progulske-Fox A., Erdos G. W., Piacentini D. A., Ayakawa G. Y., Crowley P. J., Bleiweis A. S. Construction and characterization of isogenic mutants of Streptococcus mutans deficient in major surface protein antigen P1 (I/II). Infect Immun. 1989 Nov;57(11):3306–3313. doi: 10.1128/iai.57.11.3306-3313.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moisset A., Schatz N., Lepoivre Y., Amadio S., Wachsmann D., Schöller M., Klein J. P. Conservation of salivary glycoprotein-interacting and human immunoglobulin G-cross-reactive domains of antigen I/II in oral streptococci. Infect Immun. 1994 Jan;62(1):184–193. doi: 10.1128/iai.62.1.184-193.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Munro G. H., Evans P., Todryk S., Buckett P., Kelly C. G., Lehner T. A protein fragment of streptococcal cell surface antigen I/II which prevents adhesion of Streptococcus mutans. Infect Immun. 1993 Nov;61(11):4590–4598. doi: 10.1128/iai.61.11.4590-4598.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nakai M., Okahashi N., Ohta H., Koga T. Saliva-binding region of Streptococcus mutans surface protein antigen. Infect Immun. 1993 Oct;61(10):4344–4349. doi: 10.1128/iai.61.10.4344-4349.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Okahashi N., Sasakawa C., Yoshikawa M., Hamada S., Koga T. Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries. Mol Microbiol. 1989 May;3(5):673–678. doi: 10.1111/j.1365-2958.1989.tb00215.x. [DOI] [PubMed] [Google Scholar]
  28. Russell M. W. Immunization against dental caries. Curr Opin Dent. 1992 Sep;2:72–80. [PubMed] [Google Scholar]
  29. Russell R. R. The application of molecular genetics to the microbiology of dental caries. Caries Res. 1994;28(2):69–82. doi: 10.1159/000261625. [DOI] [PubMed] [Google Scholar]
  30. Schachtele C. F., Harlander S. K., Bracke J. W., Ostrum L. C., Maltais J. A., Billings R. J. Streptococcus mutans dextransucrase: stimulation by phospholipids from human sera and oral fluids. Infect Immun. 1978 Dec;22(3):714–720. doi: 10.1128/iai.22.3.714-720.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shiroza T., Kuramitsu H. K. Construction of a model secretion system for oral streptococci. Infect Immun. 1993 Sep;61(9):3745–3755. doi: 10.1128/iai.61.9.3745-3755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shiroza T., Ueda S., Kuramitsu H. K. Sequence analysis of the gtfB gene from Streptococcus mutans. J Bacteriol. 1987 Sep;169(9):4263–4270. doi: 10.1128/jb.169.9.4263-4270.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith D. J., Taubman M. A., King W. F., Eida S., Powell J. R., Eastcott J. Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase. Infect Immun. 1994 Dec;62(12):5470–5476. doi: 10.1128/iai.62.12.5470-5476.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taubman M. A., Holmberg C. J., Smith D. J. Immunization of rats with synthetic peptide constructs from the glucan-binding or catalytic region of mutans streptococcal glucosyltransferase protects against dental caries. Infect Immun. 1995 Aug;63(8):3088–3093. doi: 10.1128/iai.63.8.3088-3093.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamashita Y., Bowen W. H., Burne R. A., Kuramitsu H. K. Role of the Streptococcus mutans gtf genes in caries induction in the specific-pathogen-free rat model. Infect Immun. 1993 Sep;61(9):3811–3817. doi: 10.1128/iai.61.9.3811-3817.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. von Eichel-Streiber C., Sauerborn M., Kuramitsu H. K. Evidence for a modular structure of the homologous repetitive C-terminal carbohydrate-binding sites of Clostridium difficile toxins and Streptococcus mutans glucosyltransferases. J Bacteriol. 1992 Oct;174(20):6707–6710. doi: 10.1128/jb.174.20.6707-6710.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

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