Four Types of Streptococcus mutans Based on Their Genetic, Antigenic and Biochemical Characteristics

Coykendall, Alan L.

doi:10.1099/00221287-83-2-327

Cited by 121 publications

(84 citation statements)

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“…The strains of serotypes c, e andfwere identical. This agrees with the finding that these three serotypes fall into a single genetically homogeneous group (Coykendall, 1974) and are known to have matching protein patterns on SDS-gel electrophoresis (Russell, 1976a). Members of serotypes d and g are also in a single genetic group, and they were found to possess a common antigen showing a line of partial identity with antigen B.…”

Section: Antigens Of Other Strainssupporting

confidence: 81%

Wall-associated Protein Antigens of Streptococcus mutans

Russell

1979

Journal of General Microbiology

231

203

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109When heat-killed whole organisms of Streptococcus mutans strain Ingbritt (serotype c ) were injected into rabbits, antibodies to at least 12 antigens were detectable by crossed immunoelectrophoresis. In contrast, when rabbits were immunized with organisms which had been subjected to extraction with the detergent sodium dodecyl sulphate (SDS), antibodies to only two protein antigens were found. These two proteins (A and B), while existing in a form apparently closely associated with peptidoglycan, could also be recovered from homogenates of whole organisms after sonication and from culture filtrates. Antigenic material was excreted throughout growth. SDS-polyacrylamide gradient gel electrophoresis showed A to have a molecular weight of 29000, while B had a molecular weight of 190000. Antigen B was purified to apparent homogeneity as judged by SDS-polyacrylamide gel electrophoresis and isoelectric focusing. All of six strains of serotype c examined produced antigen B. Strains of serotypes e and f also produce antigenically identical proteins and strains of serotypes d and g produce proteins which cross-reacted with antigen B. Antigen B was specifically precipitated by rabbit antiserum to human heart tissue. I N T R O D U C T I O NExperiments from a number of laboratories have clearly shown that immunization of animals with Streptococcus mutans or its subcellular fractions can confer protection against dental caries induced by this organism (Bowen et al., 1976), but the mechanism of this protection and the bacterial antigens involved remain unknown. For the development of an effective vaccine it is desirable to identify the antigen(s) involved in eliciting protective antibody, and the means by which protection is conferred. Identification and purification of such an antigen would aid in the understanding of the mechanism of protection, allow monitoring of vaccine preparations for efficacy, and facilitate study of its relationship with any toxic components of a crude vaccine. Of particular concern in the case of S. mutans is the demonstration of an antigenic relationship between S. mutans and human heart tissue (van de Rijn et al., 1976).An earlier report from this laboratory described successful protection of monkeys immunized with walls of S. mutans (Bowen et al., 1975). Walls treated with trypsin, on the other hand, fail to confer protection suggesting that the protective antigen might be protein in nature (Colman & Cohen, 1979). The present paper describes studies of proteins associated with S. mutans walls and the purification of one wall-associated antigen. The purified antigen is also demonstrated to be antigenically related to human heart tissue. M E T H O D R. R. B. R U S S E L Lyeast extract, 3 g; NaCI, 2 g; K2HP04, 2 g; NaHCO,, 1 g; glucose, 5 g; water, 1 1) and the chemically defined (CD) medium of Janda & Kuramitsu (1976) supplemented with 0.2% (w/v) Na,CO, (Terleckyj & Shockman, 1975). Antisera. New Zealand white rabbits (each 2 to 3 kg) were used for the preparation of antisera. Injection schedule...

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Section: Antigens Of Other Strainssupporting

confidence: 81%

Wall-associated Protein Antigens of Streptococcus mutans

Russell

1979

Journal of General Microbiology

231

203

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“…Occasional strains react strongly with neither, and they were placed in a separate serotype, f (33). Strains of all these serotypes are biochemically similar except that some group E strains do not ferment melibiose (11,30). Strains of all three serotypes share extensive DNA base sequences with strain NCTC 10449 (11,12).…”

Section: Streptococcus Mutans Clarke 1924 (8)mentioning

confidence: 81%

“…This strain was intolerant of 40% bile and 6.5% NaCl and would grow at 45°C but not at 10°C. Strain FA1 produces ammonia from arginine (11). Its DNA has 42.2 mol% G+C by thermal denaturation (9) and 41.3 mol% G+C by buoyant density in CsCl (15).…”

Section: Streptococcus Mutans Clarke 1924 (8)mentioning

confidence: 99%

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Proposal to Elevate the Subspecies of Streptococcus mutans to Species Status, Based on Their Molecular Composition

Coykendall

1977

International Journal of Systematic Bacteriology

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169

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It is proposed to confer species rank to the genetically distinct streptococci that have been considered subspecies of Streptococcus mutans Clarke: S . mutans subsp. rattus Coykendall, S. mutans subsp. cricetus Coykendall, and S . mutans subsp. sobrinus Coykendall. S , mutans is defined to exclude phenotypically similar bacteria that have deoxyribonucleic acid (DNA) guanine plup Oytosine contents appreciably different from 36 to 38 mol% and/or that do not demonstrate DNA base-sequence homology with the type strain. Streptococci that resemble S. mutans but that are quite disparate in molecular constitution are here regarded as comprising four new species: S. rattus (Coykendall) Deoxyribonucleic acid (DNA) hybridization differences in lactate dehydrogenases among (32) and the analysis of enzymes by serological three of the genospecies in respect to the enand electrophoretic methods have provided new zyme's kinetic response to pyruvate (5). He also approaches to bacterial taxonomy and fostered demonstrated electrophoretic differences in a molecular concept of species (27, 34). Thus, mannitol-and sorbitol-1-phosphate dehydroone may now define a species as a bacterium genases (4). Using serological methods, London that has a unique overall sequence of nucleo-found that aldolases of "S. mutans" were hetertide bases in its chromosome and a complement ogeneous and concluded that "S. mutans" inof enzymes characteristic of that species. Put cluded "divergent groups" of organisms (26). simply, we expect members of a species to not Heterogeneity has also been observed in polyonly "look alike" and "act alike," but also to be saccharide-synthesizing enzymes (7, 28) and in constructed of, and controlled by, molecules invertase (J. M. Tamer et al., J. Dent. Res., vol that are alike. Thus, members of a species 54, special issue A; Abstr. 53rd Gen. Session should be molecularly homologous. Bacteria Inter. Assoc. Dent. Res., 1975). S. mutans genthat are molecularly disparate should not be ospecies correlate well with the serological called by the same species name. groups described by Bratthall (2). Additional Some organisms reported to resemble Strep-serogroups found by Perch et al. (30) seem to tococcus mutans Clarke 1924 (8) are phenotypi-represent variants within the genospecies (12). cally quite similar (6,16,17,30) but are molecu-Differences have been observed in cell wall carlarly heterogeneous. Analysis of their DNA bohydrates among some genospecies (24), and base-sequence similarities has shown the exist-some morphological differences have been reence of four genetic groups (ll), which we call ported (13). "genospecies," a term introduced by Ravin (31).It is clear that "S. mutans" is actually a DNA from strains within each genospecies hy-group of streptococci, but, because of the overall bridized almost completely, but DNA from biochemical similarity of all S. mutans strains, strains of different genospecies hybridized it was proposed to conserve the nomenspecies poorly (10,ll). Furthermore, intergroup hybrid "Streptoc...

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“…The latter species appears to be very heterogeneous, and has been divided into five biotypes (Shklair & Keene, 1976), four genetic groups (Coykendall, 1974) and eight (a to h) serological groups (Bratthall, 1970;Perch et al, 1974; Beighton et al, 198 1). Since purified serotype antigens were proposed as possible vaccines against dental caries (Taubman, 1973) much attention has been given to those characterized as wall-associated polysaccharides containing, in some cases, lipoteichoic acid (Silvestri et al, 1978;Vaught & Bleiweis, 1974).…”

Section: Introductionmentioning

confidence: 99%

Protoplast and Cytoplasmic Membrane Preparations from Streptococcus sanguis and Streptococcus mutans

et al. 1983

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Protoplasts were prepared from Streptococcus sanguis and some S. mutans serotypes by use of lysozyme (EC 3.2.1.17) under particular conditions: cells had to be grown in DL-threonine (20 mM) and harvested in early exponential phase. The efficiency of protoplast formation was enhanced by two additional steps: plasmolysis (in 12% PEG), prior to addition of lysozyme, and a swirling phase, after the enzymic action. This procedure allowed us to obtain clean protoplasts, with only 0.5 % contamination by bacterial cell walls. Up to 90% protoplast lysis was obtained in 0.5 M-NaCl. Cytoplasmic membrane purification was achieved by centrifugation on a glycerol cushion. INTRODUCTIONStreptococcus sanguis and Streptococcus mutans are commonly associated with dental caries (Guggenheim, 1968). The latter species appears to be very heterogeneous, and has been divided into five biotypes (Shklair & Keene, 1976), four genetic groups (Coykendall, 1974) and eight (a to h) serological groups (Bratthall, 1970;Perch et al., 1974; Beighton et al., 198 1). Since purified serotype antigens were proposed as possible vaccines against dental caries (Taubman, 1973) much attention has been given to those characterized as wall-associated polysaccharides containing, in some cases, lipoteichoic acid (Silvestri et al., 1978;Vaught & Bleiweis, 1974). Because of the poor cross-reactivity between the different serotypes, more recent studies have dealt with the characterization of extracellular or wall-associated proteins (Russell & Lehner, 1978;Russell, 1979;Scholler et al., 1981). The difficulty in obtaining pure plasma membranes, free of cell wall components, from S. mutans and S. sanguis stems from the lack of efficient methods of protoplast preparation. Hence their cytoplasmic membranes are poorly characterized. Spheroplast induction with phage-associated lysins has been described for streptococci of Lancefield groups A, C, D and H (Krause, 1972;Bleiweis & Zimmermann, 1961 ;Calandra et al., 1975), but not for oral streptococci. Except for those belonging to Lancefield group D, most streptococci are very resistant to lysozyme lysis (Smith, 1973).Lysozyme (N-acetylmuramidase, EC 3.2.1.17) exerts its lytic effect through hydrolysis of specific glycosidic linkages in its substrate, the peptidoglycan of the bacterial cell wall. Streptococci, and especially strains of S. mutans and S. snnguis, are considered refractory to lysozyme action (Gibbons et al., 1966) and few studies have shown lysis of S. mutans cell walls with the enzyme alone (Coleman et al., 1970). Bacterial resistance to lysozyme may stem fror. several factors including the three-dimensional structure of peptidoglycan (Chiu et al., 1974 ;Formanek et al., 1974), the presence of teichoic acids covalently bound to peptidoglycan (Joseph & Shockman, 1975;Markham et al., 1975), the presence of unacetylated glucosamine residues in peptidoglycan (Hayashi et al., 1973) and surface adsorption of macromolecules from growth media (Chassy, 1976

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Four Types of Streptococcus mutans Based on Their Genetic, Antigenic and Biochemical Characteristics

Cited by 121 publications

References 6 publications

Wall-associated Protein Antigens of Streptococcus mutans

Wall-associated Protein Antigens of Streptococcus mutans

Proposal to Elevate the Subspecies of Streptococcus mutans to Species Status, Based on Their Molecular Composition

Protoplast and Cytoplasmic Membrane Preparations from Streptococcus sanguis and Streptococcus mutans

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