COMPLETE STRUCTURE OF THE CELL-SURFACE POLYSACCHARIDE OF STREPTOCOCCUS-ORALIS-C104 - A 600-MHZ NMR-STUDY

Specific lectin-carbohydrate interactions between certain oral streptococci and actinomyces contribute to the microbial colonization of teeth. The receptor molecules of Streptococcus oralis, 34, ATCC 10557, and Streptococcus mitis J22 for the galactose and N-acetylgalactosamine reactive fimbrial lectins of Actinomyces viscosus and Actinomyces naeslundii are antigenically distinct polysaccharides, each formed by a different phosphodiester-linked oligosaccharide repeating unit. These streptococci all coaggregated strongly with both A. viscosus and A. naeslundii strains, whereas S. oralis C104 interacted preferentially with certain strains of the latter species. Receptor polysaccharide was isolated from S. oralis C104 cells and was shown to contain galactose, N-acetylgalactosamine, ribitol, and phosphate with molar ratios of 4:1:1:1. The H-1 NMR spectrum of the polysaccharide shows that it contains a repeating structure. The individual sugars in the repeating unit were identified by H-1 coupling constants observed in E-COSY and DQF-COSY spectra. NMR methods included complete resonance assignments (H-1 and C-13) by various homonuclear and heteronuclear correlation experiments that utilize scalar couplings. Sequence and linkage assignments were obtained from the heteronuclear multiple-bond correlation (HMBC) spectrum. This analysis shows that the receptor polysaccharide of S. oralis C104 is a ribitol teichoic acid polymer composed of a linear hexasaccharide repeating unit containing two residues each of galactopyranose and galactofuranose and a residue each of GalNAc and ribitol joined end to end by phosphodiester linkages with the following structure. [--> 6)Gal(f)(beta-1 --> 3)Gal(p)(beta-1 --> 6)Gal(f)(beta-1 --> 6)Gal(p)NAc(beta-1 --> 3)Gal(p)(alpha-1 --> 1)ribitol(5 --> PO4-]n