Purification of dextran-binding protein from cariogenic Streptococcus mutans

McCabe, Mead M.; Hamelik, Ronald M.; Smith, Eric E.

doi:10.1016/0006-291x(77)91250-5

Cited by 58 publications

(36 citation statements)

References 19 publications

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“…McCabe et al (17) were the first to purify an extracellular GBL from S. sobrinus 6715. This protein was shown to be capable of binding specifically to ␣-1,6-linked glucans, including those synthesized by the S. sobrinus GTFs.…”

Section: Discussionmentioning

confidence: 99%

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Multiple glucan-binding proteins of Streptococcus sobrinus

Lassiter

Banas

et al. 1996

J Bacteriol

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Several proteins from culture supernatants of Streptococcus sobrinus were able to bind avidly to Sephadex G-75. The proteins could be partially eluted from the Sephadex by low-molecular-weight ␣-1,6 glucan or fully eluted by 4 M guanidine hydrochloride. Elution profiles were complex, yielding proteins of 16, 45, 58 to 60, 90, 135, and 145 kDa, showing that the wild-type strain possessed multiple glucan-binding proteins. Two mutants of Streptococcus sobrinus incapable of aggregation by high-molecular-weight ␣-1,6 glucan were isolated. One mutant was spontaneous, from a cell suspension to which glucan had been added, whereas the other was induced by ethyl methanesulfonate. Both mutants were devoid of a 60-kDa protein, as shown by gel electrophoresis of culture supernatants and whole cells. Amino acid analysis showed that the 58-to 60-kDa protein and the 90-kDa protein were distinct, although both were N-terminally blocked. Both mutants retained their ability to adhere to glass in the presence of sucrose and to ferment mannitol and sorbitol. Both mutants retained their glucosytransferase activities, as shown by activity gels. Western blots (immunoblots), employing antibody against a glucan-binding protein of Streptococcus mutans, failed to reveal cross-reactivity with S. sobrinus proteins. The results show that even though S. sobrinus produces several proteins capable of binding ␣-1,6 glucans, the 60-kDa protein is probably the lectin needed for glucan-dependent cellular aggregation.Bacterial lectins seem to have evolved for the purpose of permitting the bacteria to adhere to glycoconjugate-containing surfaces. Members of the genus Streptococcus, found in the oral cavities of humans and animals, are able to adhere to a variety of surfaces when incubated in the presence of sucrose. The disaccharide sucrose serves as a substratum for glucosyltransferases (GTFs) to yield glucans rich in ␣-1,6 and/or ␣-1,3 linkages. Many of the streptococci produce glucan-binding lectins (GBLs) or glucan-binding proteins (GBPs). Several years ago, McCabe and colleagues were able to purify streptococcal GBPs by using Sephadex as an affinity matrix (12,16,17). Others have now used similar chromatographic techniques to purify streptococcal GBPs (2,14,22,23,28,29). Banas et al. (3) were able to clone and sequence a GBP from Streptococcus mutans Ingbritt. In addition to GBLs, the streptococcal GTFs possess glucan-binding domains (19,27). Many of the bacteria which possess cell surface GTFs are not aggregated by glucans, suggesting that the GTFs do not act as lectins. The purpose of the present study was to define which surface protein of Streptococcus sobrinus is involved in glucan-dependent aggregation. To avoid confusion, a GBP is defined as any protein capable of binding reversibly to Sephadex, whereas a GBL is defined as any Sephadex-binding protein capable of conferring the ability of the bacteria to be aggregated by ␣-1,6 glucan. It is here shown that S. sobrinus possesses at least five GBPs, but only one of these, the 60-kDa band of ...

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Section: Discussionmentioning

confidence: 99%

“…Many of the streptococci produce glucan-binding lectins (GBLs) or glucan-binding proteins (GBPs). Several years ago, McCabe and colleagues were able to purify streptococcal GBPs by using Sephadex as an affinity matrix (12,16,17). Others have now used similar chromatographic techniques to purify streptococcal GBPs (2,14,22,23,28,29).…”

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confidence: 99%

Multiple glucan-binding proteins of Streptococcus sobrinus

Lassiter

Banas

et al. 1996

J Bacteriol

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“…It appears that the major role of glucans in the ecology of S. mutans is limited to enhancement of accumulation and rein forcement of the microcolony via the glucan receptors described by McCabe et al [1977] rather than interacting with the tooth sur face. If the attachment-accumulation or two reaction model of S. mutans is valid, then it is apparent w'hy the indigenous oral DP mi crobes exert little control over the coloniza tion of cariogenic microbes because dcxtranases have no effect on the initial attach ment mechanism of the cells to solid sur faces [Staat et al, 1980).…”

Section: Discussionmentioning

confidence: 99%

In vivo Relationships of the Dextran-Degrading Oral Microbiota to Streptococcus mutans and Caries Experience

1982

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Several dextranase-producing (DP) microorganisms indigenous to ths oral cavity reduce in vitro adherence of Streptococcus mutans to smooth surfaces. In this report, the DP microorganisms were studied to determine if they had an influence on the in vivo S. mutans population and the development of carious lesions in children. The epidemiological investigation showed that the oral cavity of all children sampled harbored dextranolytic microbes and that the highest concentrations were recovered from saliva. Estimates of correlation calculated from the data produced no significant antagonistic relationships between the DP bacteria found in saliva, smooth surface plaque and interproximal plaque and either the DMFS index or the S. mutans population. It was concluded that dextranase-producing microorganisms did not detectably influence the in vivo ecology of S. mutans, possibly because recent evidence indicates a nonglucan molecule may be responsible for initial attachment of S. mutans to smooth surfaces.

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“…They suggested that there are two types of dextran-binding sites on the surface of these organisms: one is cell-bound glucosyltransferase and the other undefined. Recently, dextran-binding protein was purified from an 18 hr-culture liquor of cariogenic Streptococcus mutans by affinity chromatography on Sephadex G50 (17).…”

Section: Binding Of Radioactive Glucan To Various Organismsmentioning

confidence: 99%

Mechanism of Glucan‐Induced Agglutination in *Streptococcus mutans**

Suginaka

Ebisu

Kotani

1978

Microbiology and Immunology

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The binding of radioactive glucan to Streptococcus mutatis cells, which are agglutinated by dextrans, was examined. The glucan was synthesized from sucrose by extracellular glucosyltransferases from S. mutans FA‐1 and was highly branched at C‐3 and C‐6 of D‐glucose residues, containing 17% of α(1→3)inter‐chain residues. Binding of glucan to whole cells of S. mutans OMZ‐176, which were agglutinated by addition of glucan or Dextran T2000, was irreversible and followed saturation type kinetics; saturation was achieved at approximately 110 ng of glucan per ml. About 14 ng of glucan were bound per mg of the cells at the saturated concentration. The heated cells of this organism, however, had a relatively low ability of glucan‐binding, compared with the freshly prepared and lyophilized cells. Binding to the heated cells was entirely of a non‐saturation type. Binding of Dextran T2000 or T10 was determined by competition between the labeled glucan and unlabeled Dextrans for the binding site(s). Both Dextrans and glucan from S. mutans FA‐1 were bound to the same site(s). Other organisms, which did not undergo glucan‐ and Dextran‐induced agglutination, had a relatively lower ability of glucan‐binding than S. mutans, which was agglutinated.

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Purification of dextran-binding protein from cariogenic Streptococcus mutans

Cited by 58 publications

References 19 publications

Multiple glucan-binding proteins of Streptococcus sobrinus

Multiple glucan-binding proteins of Streptococcus sobrinus

In vivo Relationships of the Dextran-Degrading Oral Microbiota to Streptococcus mutans and Caries Experience

Mechanism of Glucan‐Induced Agglutination in *Streptococcus mutans**

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Purification of dextran-binding protein from cariogenic Streptococcus mutans

Cited by 58 publications

References 19 publications

Multiple glucan-binding proteins of Streptococcus sobrinus

Multiple glucan-binding proteins of Streptococcus sobrinus

In vivo Relationships of the Dextran-Degrading Oral Microbiota to Streptococcus mutans and Caries Experience

Mechanism of Glucan‐Induced Agglutination in Streptococcus mutans*

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Mechanism of Glucan‐Induced Agglutination in *Streptococcus mutans**