Differential inhibition of Streptococcus mutans in vitro adherence by anti-glucosyltransferase antibodies

Kuramitsu, Howard K.; Ingersoll, L

doi:10.1128/iai.13.6.1775-1777.1976

Cited by 20 publications

(15 citation statements)

References 10 publications

(16 reference statements)

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“…This is supported by the fact that very little label was found on cells reacted with anti-0.2 M that had been absorbed with mutant cells. Kuramitsu and Ingersoll (37) found that antibodies directed against a high-molecular-weight enzyme fraction responsible for synthesizing primarily water-insoluble dextrans significantly inhibited in vitro adherence of S. mutans GS-5. However, antibodies against a soluble PS-syn-thesizing fraction were considerably less inhibitory.…”

Section: Resultsmentioning

confidence: 99%

Ultrastructural Localization of Sucrases in Streptococcus mutans GS-5 and an Extracellular Polysaccharide Mutant: a Comparative Cytochemical and Immunocytochemical Study

1977

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Electron microscopy and cytochemical and immunocytochemical procedures were used to study the ultrastructural distribution of sucrase enzymes in two strains of Streptococcus mutans. In a strongly adherent and virulent parent strain, GS-5, most of the invertase and fructosyltransferase activities were demonstrated extracellularly or bound to the cell surfaces. Intracellularly, enzymatic sites were detected near the plasma membrane on the periphery of the nucleoid and central mesosome. In GS-511, a mutant of diminished virulence and adherence, most of the enzymatic activity was not located on the cell surfaces, but was found away from the cell walls and associated with extracellular polysaccharides. Intracellularly, GS-511 manifested the same distribution of invertase and fructosyltransferase as did GS-5; however, the close association of these enzymes with the plasma membrane was not shown in GS-511. In both strains, extracellular areas near regions associated with cross wall formation appeared to show localized concentrations of these sucrases. Antibodies against partially purified glucosyltransferase (GTF) enzymes from GS-5 were used to localize GTF by immunocytochemical techniques. Indirect ferritin localization procedures showed that the extracellular and cell-bound GTF enzymes were distributed in similar locations as the fructosyltransferase and invertase enzymes. By absorption of the antiserum with whole GS-511 cells, the location of extracellular GTF and surface antigens unique to GS-5 was demonstrated. The dramatically reduced levels ofcell-bound sucrase activity in GS-511 indicates the significant role of these enzymes in adherence and cariogenicity. 447 on August 1, 2020 by guest http://iai.asm.org/ Downloaded from

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Section: Resultsmentioning

confidence: 99%

Ultrastructural Localization of Sucrases in Streptococcus mutans GS-5 and an Extracellular Polysaccharide Mutant: a Comparative Cytochemical and Immunocytochemical Study

1977

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“…During many studies designed to measure antibodies directed against S. mutans dextransucrase preparations, it has been consistently ob-served that control and sometimes immune sera caused an elevation rather than an inhibition of glucan production from sucrose (4,9,10,20,22,27). Based on studies with sera and oral fluids from rats, Burckhardt and Guggenheim (4) proposed that there were sites on the S. mutans dextransucrase which could interact with molecules in the fluids and enhance glucan production.…”

Section: Discussionmentioning

confidence: 99%

“…Enhanced glucan production results from an increase in the velocity of polymerization of the glucosyl moiety of sucrose due to a high affinity interaction between phospholipid and a site on the enzyme which appears to be distinct from either the glucosyl donor or glucosyl acceptor sites. We have proposed (28) that the observed stimulation of dextransucrase activity by control and/or immune sera and oral fluids from monkeys (27), rabbits (9,10,20,22), and rats (4) is due to the presence of stimulatory phospholipids in these materials.…”

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confidence: 99%

“…The methods for quantitation of total alcohol-insoluble, water-soluble, and water-insoluble glucan production from ['4C]sucrose have been described previously (12,14). Unless specified, primer dextran was present at a concentration of 20 ,M. When test components were in a solvent other than 0.01 M sodium acetate (pH 5.5), appropriate solvent controls were utilized as described elsewhere (17). Human sera.…”

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confidence: 99%

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Streptococcus mutans dextransucrase: stimulation by phospholipids from human sera and oral fluids

Schachtele¹,

Harlander²,

Bracke³

et al. 1978

Infect Immun

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Serum, gingival crevicular fluid, and parotid, submandibular, and labial minor gland saliva from four individuals stimulated glucan formation from sucrose by the Streptococcus mutans strain 6715 dextransucrase (EC 2.4.1.5). At final dilutions of 1:10 all of the fluids stimulated crude enzyme preparations approximately 1.8-fold. The fluids stimulated the purified water-insoluble glucan-synthesizing form of the dextransucrase approximately 3.2-fold and the water-soluble glucanproducing form of the enzyme approximately 2.4-fold. The fluids all contained concentrations of stimulatory material that could be reduced to undetectable levels only after dilutions of greater than 1:1,000. The increased rates of glucan formation caused by the fluids and dextran were additive, indicating that stimulation by the fluids was primarily due to interactions with entities other than glucan primer molecules. In contrast, the elevated levels of glucan formation in the presence of the fluids was not further enhanced by the addition of lysophosphatidylcholine. Lysophosphatidylcholine purified from parotid and submandibular saliva by solvent extraction and thin-layer chromatography stimulated the dextransucrase as effectively as egg yolk lysophosphatidylcholine. Thus, phospholipids normally found in human oral fluids can enhance the activity of an enzyme believed to be directly associated with the cariogenic potential of S. mutans.

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“…Recent results also suggest that the extracellular GTF activity serves as a precursor to the cellassociated form (14). In addition, both enzymatic (2,4) and immunological (19) evidence imply that more than one species of GTF may be elaborated by a single strain of S. mutans. Furthermore, characterization of the GTF activity of several different strains ofS.…”

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confidence: 99%

Interaction of glucosyltransferase with the cell surface of Streptococcus mutans

Kuramitsu¹,

Ingersoll²

1978

Infect Immun

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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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Differential inhibition of Streptococcus mutans in vitro adherence by anti-glucosyltransferase antibodies

Abstract: 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.

Cited by 20 publications

References 10 publications

Ultrastructural Localization of Sucrases in Streptococcus mutans GS-5 and an Extracellular Polysaccharide Mutant: a Comparative Cytochemical and Immunocytochemical Study

Ultrastructural Localization of Sucrases in Streptococcus mutans GS-5 and an Extracellular Polysaccharide Mutant: a Comparative Cytochemical and Immunocytochemical Study

Streptococcus mutans dextransucrase: stimulation by phospholipids from human sera and oral fluids

Interaction of glucosyltransferase with the cell surface of Streptococcus mutans

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