Characterization of the serotype e polysaccharide antigen of Streptococcus mutans (original) (raw)
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Structure of the serotype f polysaccharide antigen of Streptococcus mutans
Carbohydrate Research, 1987
The structure of the serotype f polysaccharide antigen of Streptococcus mutans was determined by methylation analysis, periodate oxidation, and partial methanolysis, and the configuration of the anomeric linkages by t3C-n.m.r. spectroscopy, indicating the trisaccharide repeating unit -+3)-a+Rhap- [cr-D-Glcp-(1+3)]-o-L-Rhap-(1 +. The structure of the backbone of the polysaccharide was confirmed by demonstrating immunological identity between the product of Smith degradation of the S. mutans serotype f antigen and the group A-variant streptococcal polysaccharide.
Structure of the group-specific polysaccharide of group E Streptococcus
Carbohydrate Research, 1985
The structure of the group-specific polysaccharide of group E Streptococcus was determined by methylation, periodate oxidation, and partial methanolysis, and the configuration of the anomeric linkages by IH-and i3C-n.m.r. spectroscopy. The trisaccharide repeating unit-+2)-cY-L-Rhap-( l--+3)-f/M-Glcp-( l-+2)]-cu-L-Rhap-( l+ was determined.
Structure of the group G Streptococcal polysaccharide
Carbohydrate Research, 1988
The structure of the group-specific polysaccharide of group G Streptococcus was determined by means of methylation analysis and selective chemical degradations. The anomeric configurations and conformations of the sugar residues were studied by 'H-and 13Gn.m.r. spectroscopy. The tetrasaccharide repeating unit,
Structure of the complex group-specific polysaccharide of group B Streptococcus
Biochemistry, 1987
The group-specific antigen was isolated from a type Ia group B streptococcal strain and is a complex polysaccharide composed of alpha-L-rhamnopyranosyl, alpha-D-galactopyranosyl, 2-acetamido-2-deoxy-beta-D-glucopyranosyl, D-glucitol, and phosphate residues. The complexity of the group B polysaccharide antigen is evident from the fact that when depolymerized by basic hydrolysis it yielded three structurally related, but nevertheless significantly different, oligosaccharides. These oligosaccharides were obtained in different molar quantities as their monophosphate esters. This evidence strongly suggests that they are linked by phosphodiester bonds in the original group B antigen. If these oligosaccharides are in fact randomly situated throughout the linear polysaccharide, then this type of heterogeneous repeating unit is unusual for a polysaccharide of bacterial origin. However, this structural arrangement of the oligosaccharides has yet to be unambiguously established because the alternate explanation of there being three different polysaccharides in the group B antigen cannot be discounted in the evidence presented here. The oligosaccharides were enzymatically dephosphorylated, and the structures of two of the three oligosaccharides are (formula: see text) Despite their structural differences, the two oligosaccharides are related by the smaller being an integral part of the larger. In the structural analysis of the group B antigen, methylation analysis, periodate oxidation, nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, fast atom bombardment mass spectrometry, and various specific chemical and enzymatic degradations were the principal methods used. Of particular interest was the use of an alpha-rhamnosidase to selectively degrade the larger oligosaccharide. This facilitated the assignment of signals in its 1H and 13C NMR spectra.
Characterization of the group-specific polysaccharide of group B Streptococcus
Archives of Biochemistry and Biophysics, 1984
The group-specific polysaccharide of the group B Streptococcus was isolated by nitrous acid extraction followed by gel filtration on Sepharose 6B and chromatography on DEAE-Bio-Gel A. It was composed of rhamnose, galactose, N-acetylglucosamine, and glucitol phosphate. Mild periodate oxidation of the polysaccharide resulted in a rapid reduction in molecular weight, indicating that the glucitol was located in the backbone of the polymer. High-resolution 31P NMR showed the presence of a single type of phosphodiester bond in the molecule. Methylation analysis and several specific chemical degradations were done to determine sugar linkages. The basic structure of the group B polysaccharide consists of a backbone of 2-linked rhamnose, 2,4-linked rhamnose, and glucitol phosphate, and side chains of rhamnose(1 -3)galactose(l -3)N-acetylglucosamine linked to the 4-position of a rhamnose in the backbone. o
Journal of Immunological Methods, 1995
A method for the immunoblot analysis of the type-specific capsular polysaccharides of streptococci was developed. The capsular polysaccharides were extracted by sonication and subjected to polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulphate (SDS). After electrophoresis the polysaccharides were transferred to charged PVDF-N membranes and probed with the type specific antibodies. A characteristic ladder-like pattern of polysaccharide bands specific for the serotype (1, 2, 4, 7) was observed for capsular extracts of Streptococcus suis. Human immune sera against type-specific group B streptococcal polysaccharides reacted most strongly with the immunizing polysaccharide type (Ia, II, III). The previously observed crossreactions between the group B streptococcal type-specific capsular preparations were shown to be due to binding to the isomeric polysaccharide molecules. Thus, gel electrophoresis combined with immunoblot analysis seems to provide a novel method for the molecular and immunochemical characterization of bacterial polysaccharides and for the study of the specificity and properties of antibodies to capsular polysaccharides.
Journal of Biological Chemistry, 1991
The type V capsular polysaccharide of group B Streptococcus has been isolated and purified, and its repeating unit structure determined. The native type v polysaccharide contains D-glUCOSe, D-galactose, 2acetamido-2-deoxy-~-glucose, and sialic acid in a molar ratio of 3:2:1:1. Methylation analysis and 'H NMR and '"C NMR analysis of the native type V polysaccharide and of its specifically degraded products permitted the determination of the repeating unit structure of the type V polysaccharide: ~4)-a-~-G l c p-(1~4)-P-~-G a l p-(l~4)-~-D-G l c p-(l~ 6 3
Structure determination of Streptococcus suis serotype 2 capsular polysaccharide
Biochemistry and Cell Biology-biochimie Et Biologie Cellulaire, 2010
The capsular polysaccharide (CPS) of Streptococcus suis serotype 14 was purified, chemically modified, and characterized. Sugar and absolute configuration analyses gave the following CPS composition: D-Gal, 3; D-Glc, 1; D-GlcNAc, 1; D-Neu5Ac, 1. The Sambucus nigra lectin, which recognizes the Neu5Ac(␣2-6)Gal/GalNAc sequence, showed binding to the native CPS. Sialic acid was found to be terminal, and the CPS was quantitatively desialylated by mild acid hydrolysis. It was also submitted to periodate oxidation followed by borohydride reduction and Smith degradation. Sugar and methylation analyses, 1 H and 13 C nuclear magnetic resonance, and mass spectrometry of the native CPS or of its specifically modified products allowed to determine the repeating unit sequence:
Microbiological Research, 1997
A Monoclonal antibody (MAb II-T) specific for serotypes II and V of Group E streptococci (GES) was prepared by fusing myeloma cells with spleen cells of mice immunized with whole cells of a serotype II strain. MAb II -T reacted in an enzyme immunoassay (EIA) with whole cells of both serotypes and reacted in gel diffusion test with autoclaved-saline extracts of serotypes II and V. The extract was purified by DEAE-Sephadex A-25, followed by treatment with proteinase K, and further by chromatogniphy with a Sephadex G-200 column. The purified polysaccharide (PS) antigen contained 98.6% carbohydrate and 1.4% protein, but no detectable phosphorus. In hapten inhibition tests using various sugars, D-mannosamine markedly inhibited the precipitin reaction. These results indicated that the antigenic determinant might have a structure similar to D-mannosamine.
Isolation and characterization of type IV group B Streptococcus capsular polysaccharide
Infection and Immunity, 1989
An antigenically distinct serotype, type IV, has recently been added to the recognized serotypes of group B streptococci (GBS). We isolated and purified the capsular polysaccharide antigen from a prototype type IV GBS strain. The type IV capsular polysaccharide formed a precipitin line with rabbit antiserum to type IV GBS organisms but not with antiserum to organisms of GBS serotype Ia, Ib, II, or III. Enzyme-linked immunosorbent assay inhibition experiments showed no cross-reaction between type IV antiserum and other GBS serotypes. Capsular polysaccharide released from the bacterial cells with mutanolysin and that isolated from the culture supernatant had similar elution profiles on Sepharose CL-6B, with a Kav of 0.30 and an estimated Mr of 200,000. The purified type IV polysaccharide was found to contain galactose, glucose, N-acetylglucosamine, and N-acetylneuraminic acid (sialic acid) as exclusive sugars. The polysaccharide contained 23% (by weight) sialic acid and galactose, glu...