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Volume 144, Issue 5

Identification and analysis of a gene () encoding a major amylase-binding protein in Free

Abstract

Oral streptococci such as bind the abundant salivary enzyme -amylase. This interaction may be important in dental plaque formation and metabolism, thus contributing to the initiation and progression of dental caries and periodontal disease, the two most common plaque-mediated diseases. The conjugative transposon Tn was used to insertionally inactivate gene(s) essential to the expression of amylase-binding components of Challis, and a mutant deficient in amylase-binding (Challis Tn1) was identified. While wild-type strains of released both 20 kDa and 82 kDa amylase-binding proteins into culture supernatants, Challis Tn1 expressed the 82 kDa but not the 20 kDa protein. The 20 kDa amylase-binding protein was isolated from culture supernatants of Challis by hydroxyapatite chromatography. A partially purified, functionally active 20 kDa protein was sequenced from blots, and the N-terminal sequence obtained was found to be DEP (A) TDAAT(R)NND. A novel strategy, based on the single-specific-primer polymerase chain reaction technique, enabled the gene inactivated by Tn to be cloned. Analysis of the resultant nucleotide sequence revealed an open reading frame of 585 bp, designated amylase-binding protein A (), encoding a protein of 20 kDa (AbpA), immediately downstream from the insertion site of Tn This protein possessed a potential signal peptide followed by a region having identity with the N-terminal sequence of the 20 kDa amylase-binding protein. These results demonstrate the role of the 20 kDa protein in the binding of amylase to Knowledge of the nature of amylase-binding proteins may provide a better understanding of the role of these proteins in the colonization of in the oral cavity.

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Keyword(s): amylase-binding protein , biofilms , dental plaque , microbial adhesion/adherence and salivary proteins
© Society for General Microbiology 1998
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1998-05-01
2024-04-18
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References

  1. Aguirre, A., Levine, M. J., Cohen, R. E., Tabak, L. A. (1987); Immunochemical quantitation of a-amylase and secretory IgA in parotid saliva from people of various ages.. Archives of Oral Biology 32:(4)297–301 [View Article]
     [Google Scholar]
  2. Altschui, S. F., Gish, W., Miller, W., Myers, E. W., Lipman, D. J. (1990); Basic local alignment search tool.. Journal of Molecular Biology 215:(3)403–410 [View Article]
     [Google Scholar]
  3. Bairoch, A., Bucher, P., Hofmann, K. (1996); The prosite database, its status in 1995.. Nucleic Acids Research 24:(1)189–196 [View Article]
     [Google Scholar]
  4. Behnke, D. (1981); Plasmid transformation of Streptococcus sanguis (Challis) occurs by circular and linear molecules.. Molecular & General Genetics 182:(3)490–497 [View Article]
     [Google Scholar]
  5. Birnboim, H. C., Doly, J. (1979); A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Research 7:(6)1513–1523 [View Article]
     [Google Scholar]
  6. Brendel, V., Bucher, P., Nourbakhsh, I., Blaisdell, B. E., Karlin, S. (1992); Methods and algorithms for statistical analysis of protein sequences.. Proc Natl Acad Sci USA 89:(6)2002–2006 [View Article]
     [Google Scholar]
  7. Burnette, W. N. (1981); Western blotting: electrophoretic transfer of proteins from sodium dodecyl sulphate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.. Anal Biocbem 112:(2)195–203 [View Article]
     [Google Scholar]
  8. Caparon, M. G., Scott, J. R. (1987); Identification of a gene that regulates expression of M protein, the major virulence determinant of group A streptococci.. Proc Natl Acad Sci USA 84:(23)8677–8681 [View Article]
     [Google Scholar]
  9. Caufield, P. W., Shah, G. R., Hollingshead, S. K. (1990); Use of transposon Tn916 to inactivate and isolate a mutacin-associated gene from Streptococcus mutans.. Infection and Immunity 58:(12)4126–4135 [View Article]
     [Google Scholar]
  10. Cimasoni, G., Song, M., McBride, B. C. (1987); Effect of crevicular fluid and lysosomal enzymes on the adherence of streptococci and bacteroides to hydroxyapatite.. Infection and Immunity 55:(6)1484–1489 [View Article]
     [Google Scholar]
  11. Clewell, D. B. (1981); Plasmids, drug resistance, and gene transfer in the genus Streptococcus.. Microbiological Reviews 45:(3)409–436 [View Article]
     [Google Scholar]
  12. Coykendail, A. L., Gustafson, K. B. (1985); Deoxyribonucleic acid hybridizations among strains of Streptococcus salivarius and Streptococcus bovis.. International Journal of Systematic Bacteriology 35:(3)274–280 [View Article]
     [Google Scholar]
  13. Curby, W. A. (1953); Device for collection of parotid saliva.. Journal of Laboratory and Clinical Medicine 41:493–496
     [Google Scholar]
  14. Douglas, C. W. I. (1983); The binding of human salivary a-amylase by oral strains of streptococcal bacteria.. Archives of Oral Biology 28:(7)567–573 [View Article]
     [Google Scholar]
  15. Douglas, C. W. I. (1990); Characterization of the a-amylase receptor of Streptococcus gordonii NCTC 7868.. Journal of Dental Research 69:(11)1746–1752 [View Article]
     [Google Scholar]
  16. Douglas, C. W. I. (1994); Bacterial-protein interactions in the oral cavity.. Adv Dent Res 8:(2)254–262 [View Article]
     [Google Scholar]
  17. Douglas, C. W. I., Pease, A. A., Whiley, R. A. (1990); Amylasebinding as a discriminator among oral streptococci.. Fems Microbiology Letters 66:(1–3)193–198 [View Article]
     [Google Scholar]
  18. Douglas, C. W. I., Heath, J., Gwynn, J. P. (1992); Enzymic activity of salivary amylase when bound to the surface of oral streptococci.. Fems Microbiology Letters 92:(2)193–198 [View Article]
     [Google Scholar]
  19. Gawron-Burke, C., Clewell, D. B. (1984); Regeneration of insertionally inactivated streptococcal DNA fragments after excision of transposon Tn916 in Escherichia coli: strategy for targeting and cloning genes from Gram-positive bacteria.. Journal of Bacteriology 159:(1)214–221 [View Article]
     [Google Scholar]
  20. Giulian, G. G., Moss, R. L., Greaser, M. (1983); Improved methodology for analysis and quantitation of proteins on onedimensional silver-stained slab gels.. Analytical Biochemistry 129:(2)277–287 [View Article]
     [Google Scholar]
  21. Gwynn, J. P. (1995); Characterisation of the interactions between human salivary a-amylase and oral streptococci,. PhD thesis. University of Sheffield;
  22. Gwynn, J. P., Douglas, C. W. I. (1994); Comparison of amylasebinding proteins in oral streptococci.. Fems Microbiology Letters 124:(3)373–380 [View Article]
     [Google Scholar]
  23. Handley, P., Coykendail, A., Beighton, D., Hardie, J. M., Whiley, R. A. (1991); Streptococcus crista sp. nov., a viridans streptococcus with tufted fibrils, isolated from the human oral cavity and throat.. International Journal of Systematic Bacteriology 41:(4)543–547 [View Article]
     [Google Scholar]
  24. Harris, G. S., Michalek, S. M., Curtis, R. C. Ill (1992); Cloning of a locus involved in Streptococcus mutans intracellular polysaccharide accumulation and virulence testing of an intracellular polysaccharide-deficient mutant.. Infection and Immunity 60:(8)3175–3185 [View Article]
     [Google Scholar]
  25. Higgins, M. L., Shockman, G. D. (1970); Model for cell wall growth of Streptococcus faecalis.. Journal of Bacteriology 101:(2)643–648 [View Article]
     [Google Scholar]
  26. Jenkinson, H. F. (1994); Cell surface protein receptors in oral streptococci.. Fems Microbiology Letters 121:(2)133–140 [View Article]
     [Google Scholar]
  27. Kilian, M., Nyvad, B. (1990); Ability to bind salivary a-amylase discriminates certain viridans streptococcal species.. Journal of Clinical Microbiology 28:(11)2576–2577 [View Article]
     [Google Scholar]
  28. Laemmli, U. K. (1970); Cleavage of structural proteins during assembly of the head of bacteriophage T4.. Nature 277:(5259)680–685 [View Article]
     [Google Scholar]
  29. LeBlanc, D. J., Hassell F. P. (1976); Transformation of Streptococcus sanguis Challis by plasmid deoxyribonuceic acid from Streptococcus faecalis.. ] Bacteriol 128:(1)347–355 [View Article]
     [Google Scholar]
  30. Lottenberg, R., Broder, C. C., Boyle, M. D. P., Kain, S. J., Schroeder, B. L., Curtiss R. (1992); Cloning, sequence analysis and expression in Escherichia coli of a streptococcal plasmin receptor.. Journal of Bacteriology 174:(16)5240–5210 [View Article]
     [Google Scholar]
  31. Macrina, F. L., Evans, R. P., Tobian J.A., Hartley, D. L., Clewell, D. B., Jones K. R. (1983); Novel shuttle plasmid vehicles for Escherichia-Streptococcus transgeneric cloning.. Gene 25:145159
     [Google Scholar]
  32. Merril, C. R., Goldman, D., van Keuren, M. L. (1982); Simplified silver protein detection and image enhancement methods in polyacrylamide gels.. Electrophoresis 3:(1)17–23 [View Article]
     [Google Scholar]
  33. Novak, J., Novak, L., Shah, G. R., Woodruff, W. A., Caufield P. W. (1997); Transposon mutagenesis, cloning of chromosomal DNA from the site of Tn916 insertion using polymerase chain reaction.. Biotechnology Techniques 11:(1)51–54 [View Article]
     [Google Scholar]
  34. Roger, V., Tenovuo, J., Lenander-Lumikari, M., Soderling, E., Vilja, P. (1994); Lysozyme and lactoperoxidase inhibit the adherence of Streptococcus mutans NCTC 10449 (serotype c) to saliva-treated hydroxyapatite in vitro.. Caries Research 28:(6)421–428 [View Article]
     [Google Scholar]
  35. Sanger, F., Nicklen, S., Coulson A. R. (1977); DNA sequencing with chain terminating inhibitors.. Proc Nat Acad Sci USA 74:(12)5463–5467 [View Article]
     [Google Scholar]
  36. Scannapieco, F. A. (1994); Saliva-bacterium interactions in oral microbial ecology.. Crit Rev Oral Biol Med 5:(3)203–248 [View Article]
     [Google Scholar]
  37. Scannapieco, F. A., Bergey, E. J., Reddy M.S., Levine, M. J. (1989); Characterization of salivary a-amylase binding to Streptococcus sanguis.. Infection and Immunity 57:(9)2853–2863 [View Article]
     [Google Scholar]
  38. Scannapieco, F. A., Haraszthy, G. G., Cho, M. I., Levine M. J. (1992); Characterization of an amylase-binding component from Streptococcus gordonii G9B.. Infection and Immunity 60:(11)4726–4733 [View Article]
     [Google Scholar]
  39. Scannapieco, F. A., Solomon, L., Wadenya, R. O. (1994); Emergence in human dental plaque and host distribution of amylase-binding streptococci.. Journal of Dental Research 73:(10)1627–1635 [View Article]
     [Google Scholar]
  40. Scannapieco, F. A., Torres, G. I., Levine, M. J. (1995); Salivary amylase promotes adhesion of oral streptococci to hydroxyapatite.. Journal of Dental Research 74:(7)1360–1366 [View Article]
     [Google Scholar]
  41. Schneewind, 0., Fowler, A., Faull, K. F. (1995); Structure of the cell wall anchor of surface proteins in Staphylococcus aureus.. Science 268:(5207)103–106 [View Article]
     [Google Scholar]
  42. Shyamala, V., Ames, G. F. (1989); Genome walking by singlespecific-primer polymerase chain reaction: SSP-PCR.. Gene 84:(1)1–8 [View Article]
     [Google Scholar]
  43. Sutcliffe, I. C., Russell, R. R. (1995); Lipoproteins of Grampositive bacteria.. J Bacterial T1:(5)1123–1128 [View Article]
     [Google Scholar]
  44. Suvorov, A. N., Flores, A. E., Ferrieri, P. (1997); Cloning of the glutamine synthetase gene from group B streptococci.. Infection and Immunity 65:(1)191–196 [View Article]
     [Google Scholar]
  45. Tellefson, L. M., Germaine, G. R. (1986); Adherence of Streptococcus sanguis to hydroxyapatite coated with lysozyme and lysozyme-supplemented saliva.. Infection and Immunity 51:(3)750–759 [View Article]
     [Google Scholar]
  46. Terleckyj, B., Willet, N. P., Schockman G. D. (1975); Growth of several cariogenic strains of oral streptococci in a chemically defined medium.. Infection and Immunity 11:(4)649–655 [View Article]
     [Google Scholar]
  47. Tseng, C. C., Scannapieco, F. A., Levine, M. J. (1992); Use of a replica plate assay for the rapid assessment of salivary protein- bacteria interactions.. Oral Microbiol Immunol 7:(1)53–56 [View Article]
     [Google Scholar]
  48. Yamamoto, M., Jones, J. M., Senghas, E., Gawron-Burke, C., Clewell, D. B. (1987); Generation of Tn5 insertions in streptococcal conjugative transposon Tn916.. Appl Environ Microbiol 53:(5)1069–1072 [View Article]
     [Google Scholar]
  49. Yanisch-Perron, C., Vieira, J., Messing, J. (1985); Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mpl8 and pUC19 vectors.. Gene 33:(1)103–119 [View Article]
     [Google Scholar]
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Identification and analysis of a gene (abpA) encoding a major amylase-binding protein in Streptococcus gordonii
Microbiology 144, 1223 (1998); https://doi.org/10.1099/00221287-144-5-1223
/content/journal/micro/10.1099/00221287-144-5-1223
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