|
Sign In to gain access to subscriptions and/or personal tools.
|
Characterization of GlucosyltransferaseB, GtfC, and GtfD in Solution and on the Surface of Hydroxyapatite
A.R. Venkitaraman
Department of Dental Research, Box 611, Rochester Caries Research Center, University of Rochester Medical Center, Rochester, New York 14642
A.M. Vacca-Smith
Department of Dental Research, Box 611, Rochester Caries Research Center, University of Rochester Medical Center, Rochester, New York 14642
L.K. Kopec
Department of Dental Research, Box 611, Rochester Caries Research Center, University of Rochester Medical Center, Rochester, New York 14642
W.H. Bowen
Department of Dental Research, Box 611, Rochester Caries Research Center, University of Rochester Medical Center, Rochester, New York 14642
GlucosyltransferaseB, GtfC, and GtfD were purified by hydroxyapatite column chromatography, followed by ultrafiltration from the culture supernatant fluids of three Streptococcus milleri constructs (gift from Dr. H.K. Kuramitsu) which harbored individual gtf genes of Streptococcus mutans GS5. GtfB, GtfC, and GtfD were enzymatically active both in solution and in an experimental pellicle (HA-CWS-Gtf) formed by adsorbing Gtf onto the surface of clarified human whole saliva (CWS)-coated hydroxyapatite (HA). The Km values for sucrose for all three enzymes were lower when the enzyme was adsorbed to a surface, compared with when it was in solution. In solution phase assays, and in the absence of primer dextran, glucan production was enhanced 75% when both GtfB and GtfD were present in the reaction mixture, compared with the sum of the individual enzyme activities (p < 0.005). This enhancement did not occur when GtfC was additionally present, or when the GtfB + GtfD enzyme pair was adsorbed onto HA-CWS. In additional experiments, glucan formed by GtfB or GtfC, but not by GtfD, on a HA-CWS-Gtf surface increased adherence of Streptococcus mutans GS5 and Streptococcus sobrinus 6715 by seven- to nine-fold compared with adherence when no glucan was present on the pellicle surface (p < 0.001). Further, treatment of the HA-CWS-GtfB-glucan or HA-CWS-GtfC-glucan pellicle with a-1,6 dextranase significantly reduced adherence of both streptococcal strains (p < 0.001). These results show that GtfB, GtfC, and GtfD are enzymatically active in an adsorbed state and that the nature of their product glucan can influence the adherence of cariogenic oral streptococci to an experimental pellicle.
Key Words: glucosyltransferases • Streptococcus mutans • pellicle • adherence, plaque
REFERENCES
- Aoki H., Shiroza T., Hayakawa M., Sato S., Kuramitsu HK (1986). Cloning of a Streptococcus mutans glucosyltransferase gene coding for insoluble glucan synthesis. Infect Immun 53:587-594. Bailey JE, Ollis DF (1986). Biochemical engineering fundamentals. 2nd ed. New York: McGraw-Hill.[Abstract/Free Full Text]
- Clark WB, Bamman LL, Gibbons RJ (1978). Comparative estimates of bacterial affinities and adsorption sites on hydroxyapatite surfaces. Infect Immun 19:846-853.[Abstract/Free Full Text]
- Douglas CWI, Russell RRB (1982). Effect of specific antisera on adherence properties of the oral bacterium Streptococcus mutans. Arch Oral Biol 27:1039-1045.[CrossRef][Medline]
[Order article via Infotrieve]
- Doyle RJ, Ciardi JE, editors (1983). Glucosyltransferases, glucans, sucrose and dental caries In: Chemical senses (spec suppl). Washington: IRL Press.
- Drake D., Taylor KG, Bleiweis AS, Doyle RJ (1988). Specificity of the glucan-binding lectin of Streptococcus cricetus. Infect Immun 56:1864-1872.[Abstract/Free Full Text]
- Fukushima K., Ikeda T., Kuramitsu HK (1992). Expression of Streptococcus mutans glucosyltransferase genes in Streptococcus milleri. Infect Immun 60:2815-2822.[Abstract/Free Full Text]
- Germaine GR, Schachtele CF, Chludzinski AM (1974). Rapid filter paper assay for the dextransucrase activity from Streptococcus mutans. J Dent Res 53:1355-1360.
- Gibbons RJ, Cohen L., Hay DI (1986). Strains of Streptococcus mutans and Streptococcus sobrinus attach to different pellicle receptors. Infect Immun 52:555-561.[Abstract/Free Full Text]
- Gilmore KS, Russell RRB, Ferretti JJ (1993). Expression of GtfS is essential for normal insoluble glucan synthesis by Streptococcus downei. Infect Immun 61:1246-1250.[Abstract/Free Full Text]
- Hamada S., Slade HD (1980). Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev 44:331-384. Hanada N., Kuramitsu HK (1988). Isolation and characterization of the Streptococcus mutans gtfc gene, coding for synthesis of both soluble and insoluble glucans. Infect Immun 56:1999-2005.[Abstract/Free Full Text]
- Hanada N., Kuramitsu HK (1989). Isolation and characterization of the Streptococcus mutans gtfd gene, coding for primer-dependent soluble glucan synthesis. Infect Immun 57:2079-2085.[Abstract/Free Full Text]
- Inoue M., Shibata H., Morioka T. (1982). Serotype specificity for
- Kelstrup J., Funder-Nielsen TD (1974). Adhesion of dextran to Streptococcus mutans. J Gen Microbiol 81:485-489.[Abstract/Free Full Text]
- Koga T., Sato S., Inoue M., Takeuchi K., Furuta T., Hamada S. (1983). Role of primers in glucan synthesis by glucosyltransferases from Streptococcus mutans strain OMZ176. J Gen Microbiol 129:751-754.[Abstract/Free Full Text]
- Kuramitsu H., Smorawinska M., Yamashita Y. (1994). Molecular biology of Streptococcus mutans virulence. In: Cariology for the nineties. Bowen WH, Tabak LA, editors. Rochester: University of Rochester Press, pp. 301-308.
- Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685. Lee SF, Progulske-Fox A., Bleiweis AS (1988). Molecular cloning and expression of a Streptococcus mutans major surface protein antigen, P1 (I/II), in Escherichia coli. Infect Immun 56:2114-2119.[Abstract/Free Full Text]
- Loesche WJ (1986). Role of Streptococcus mutans in human dental decay. Microbiol Rev 50:353-380.[Free Full Text]
- McCabe MM (1985). Purification and characterization of a primer-independent glucosyltransferase from Streptococcus mutans 6715-13 mutant 27. Infect Immun 50:771-777. McCormack SM, Tormo FJ, Featherstone JDB (1991). A straightforward scanning electron microscopy technique for examining non-metal coated dental hard tissues. Scanning Microsc 5:269-272.
- Morrissey JH (1981). Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307-310.[CrossRef][Medline]
[Order article via Infotrieve]
- Munro C., Michalek SM, Macrina FL (1991). Cariogenicity of Streptococcus mutans V403 glucosyltransferase and fructosyltransferase mutants constructed by allelic exchange. Infect Immun 59:2316-2323.[Abstract/Free Full Text]
- Ohta H., Kato H., Okahashi N., Takahashi I., Hamada S., Koga T. ( 1989). Characterization of a cell-surface protein antigen of hydrophilic Streptococcus mutans strain GS-5. J Gen Microbiol 135:981-988.[Abstract/Free Full Text]
- Robrish SA, Reid W., Krichevsky MI (1972). Distribution of enzymes forming polysaccharide from sucrose and the composition of extracellular polysaccharide synthesized by Streptococcus mutans. Appl Micobiol 24:184-190.[Medline]
[Order article via Infotrieve]
- Russell RRB (1994). The application of molecular genetics to the microbiology of dental caries. Caries Res 28:69-82. Schilling KM, Bowen WH (1988). The activity of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite. J Dent Res 67:2-8.
- Schilling KM, Bowen WH (1992). Glucan synthesized in situ in experimental salivary pellicle functions as specific binding sites for Streptococcus mutans. Infect Immun 60:284-295. Wu-Yuan CD, Tai S, Slade HD (1979). Properties of Streptococcus mutans grown in a synthetic medium: binding of glucosyltransferase and in-vitro adherence and binding of dextran-glucan and glycoprotein and agglutination. Infect Immun 23:600-608.
- Yamashita Y., Bowen WH, Burne RA, Kuramitsu HK (1993). Role of the Streptococcus mutans glucosyltransferase genes in caries induction in the specific-pathogen-free rat model. Infect Immun 61:3811-3817.[Abstract/Free Full Text]
Journal of Dental Research, Vol. 74, No. 10,
1695-1701 (1995)
DOI: 10.1177/00220345950740101101
 CiteULike  Complore  Connotea  Del.icio.us  Digg  Reddit  Technorati  Twitter What's this?
This article has been cited by other articles:
|
|
|
|
 
M. I. Klein, S. Duarte, J. Xiao, S. Mitra, T. H. Foster, and H. Koo
Structural and Molecular Basis of the Role of Starch and Sucrose in Streptococcus mutans Biofilm Development
Appl. Envir. Microbiol.,
February 1, 2009;
75(3):
837 - 841.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S.-J. Ahn, Z. T. Wen, and R. A. Burne
Effects of Oxygen on Virulence Traits of Streptococcus mutans
J. Bacteriol.,
December 1, 2007;
189(23):
8519 - 8527.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
X. Shi, N. N. Rao, and A. Kornberg
Inorganic polyphosphate in Bacillus cereus: Motility, biofilm formation, and sporulation
PNAS,
December 7, 2004;
101(49):
17061 - 17065.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Koo, M. F. Hayacibara, B. D. Schobel, J. A. Cury, P. L. Rosalen, Y. K. Park, A. M. Vacca-Smith, and W. H. Bowen
Inhibition of Streptococcus mutans biofilm accumulation and polysaccharide production by apigenin and tt-farnesol
J. Antimicrob. Chemother.,
November 1, 2003;
52(5):
782 - 789.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Koo, P. L. Rosalen, J. A. Cury, Y. K. Park, and W. H. Bowen
Effects of Compounds Found in Propolis on Streptococcus mutans Growth and on Glucosyltransferase Activity
Antimicrob. Agents Chemother.,
May 1, 2002;
46(5):
1302 - 1309.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. H. Bowen
DO WE NEED TO BE CONCERNED ABOUT DENTAL CARIES IN THE COMING MILLENNIUM?
Critical Reviews in Oral Biology & Medicine,
March 1, 2002;
13(2):
126 - 131.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
