Antibodies prepared against an insoluble-soluble glucan-synthesizing fraction significantly inhibited in vitro adherence of Streptococcus mutans, whereas antibodies directed against a soluble glucan-synthesizing fraction were much less inhibitory.
Partially purified glucosyltransferase enzymes from Streptococcus mutans GS-5 (serotype c) have been utilized to prepare antibodies directed against the soluble glucan-synthesizing activity, GTF-B, and the insoluble-soluble glucan synthetic activity, GTF-A. Anti-GTF-A inhibited insoluble glucan formation catalyzed by the extracellular enzymes from strains GS-5 and FA-1 (serotype b) to a much greater extent than that of strain HS-6 (serotype a) or OMZ-176 (serotype d). This antibody fraction also inhibited both the cell-associated glucosyltransferase activities as well as the sucrose-mediated adherence of cells to glass surfaces by strains GS-5 and FA-1 but not that of strains HS-6 and OMZ-176. Anti-GTF-B inhibited soluble glucan formation catalyzed by the extracellular enzymes of strain GS-5 but not that of strain HS-6, FA-1, or OMZ-176. However, this antibody fraction did not strongly inhibit either the cell-associated glucosyltransferase activity or cellular adherence of any ofthe four strains. These results with both antibody fractions were also correlated with the ability of the antibodies to agglutinate the cells and form precipitin bands after immunodiffusion with the extracellular enzymes. Antibody prepared against the homogeneous soluble glucan-synthesizing enzyme demonstrated similar effects to the anti-GTF-B fraction. These results are discussed in terms ofthe antigenic relationships existing between the glucosyltransferases from different serotypes of S. mutans.
The enzymatic and adherence properties of Streptococcus mutans GS5 and S. sanguis ST3, both isolated from human carious lesions, have been compared. During growth in sucrose media, S. mutans GS5 adheres to smooth surfaces approximately three times more effectively than does S. sanguis ST3. However, strain ST3 does not display sucrose-dependent adherence under nongrowth conditions, whereas strain GS5 displays significant adherence. Although both organisms synthesize both water-soluble and -insoluble glucans, the glucosyltransferases from S. mutans GS5 synthesize much more adherent glucan molecules than do the comparable enzymes from S. sanguis ST3. Both cell types bind exogenous glucosyltransferases synthesized by strain ST3 equally well, whereas cells of strain GS5 bind the comparable enzyme fraction that it synthesizes to a greater degree than do cells of S. sanguis ST3. However, in contrast to the results with cells of S. mutans GS5, the absorption of the glucosyltransferase activity synthesized by S. mutans GS5 to the surface ofS. sanguis ST3 results in low levels of subsequent sucrose-dependent adherence. These results are discussed in terms of the molecular basis for the sucrose-dependent adherence of the oral streptococci to smooth surfaces.
The partially purified glucosyltransferase (GTF) fraction synthesizing primarily water-insoluble glucans, GTF-A, and the homogeneous fraction synthesizing water-soluble glucans, GTF-B, were utilized to assess the binding of GTF activity to the cell surface of Streptococcus mutans GS-5. Growth of the cells in either Todd-Hewitt broth or a chemically defined medium did not appear to affect the ability of the cells to bind either enzyme fraction. Heat inactivation of the cells did not singificantly reduce the interaction of the enzymes with the cells. Cell surface glucan molecules appear to be involved in GTF binding to the cells because: (i) dextranase or alpha-1,3-glucanase treatment of the cells markedly reduced enzyme binding; (ii) the inclusion of soluble dextrans in the binding assays reduced both GTF-A and GTF-B binding to the cells; and (iii) pretreatment of the cells or the GTF-B fraction with soluble dextrans before binding significantly reduced enzyme binding to the cells. In addition, enzyme binding appears to require a cell surface protein component because Pronase, but not trypsin, treatment of cells reduced enzyme binding. Furthermore, the removal of a portion of the cell surface GTF-glucan complex with 3 N NaCl appears to provide additional binding sites for the enzymes. These results are interpreted in terms of the mechanism of the conversion of extracellular GTF to the cell-associated form.
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