Structural analysis of the surface polysaccharide of Staphylococcus aureus M

Abstract

The chemical structure of the surface polysaccharide from Staphylococcus aureus M was investigated by a combination of methanolytic, hydrolytic, and chromatographic techniques. The repeating unit that was most consistent with the data was a hexasaccharide composed of N-acetyl-D-aminogalacturonic acid, N-acetyl-D-fucosamine, and taurine in molar ratios of 4:2:1. A disaccharide was isolated and characterized, by combined gas-liquid chromatography-mass spectrometry, as N-acetyl-D-aminogalacturonyl-(1-+ 3)-N-acetyl-D-fucosamine. Taurine is linked to a carboxyl group of N-acetyl-n-aminogalacturonic acid via an amide bond. We have previously reported the isolation and identification of the constituent components of the surface polysaccharide ofStaphylococcus aureus M (2). This polysaccharide is composed of D-aminogalacturonic acid, D-fucosamine, and taurine. It thus differs from the surface polysaccharides of the S. aureus Smith diffuse variant, which is composed of D-aminoglucuronic acid and L-alanine (1), and S. aureus mutant T, which is composed of D-aminomannuronic acid and D-fucosamine (11). This report is concerned with structural analysis of the strain M surface polysaccharide. (This work was presented in part at the 59th Annual Meeting of the Federation of American Societies for Experimental Biology, Atlantic City, N.J., 1975.) MATERIALS AND METHODS Unless otherwise indicated, the materials and methods for the isolation and identification of the components of the surface polysaccharide antigen (SPA) from S. aureus M are the same as previously described (2). Other materials used in this study were obtained from the following sources: AG50W-X8 resins, Bio-Rad Laboratories; thin-layer chromatographic (TLC) plates (no. 6064 cellulose, no. 6060 silica gel with fluorescent indicator), Eastman Kodak Co.; coiled glass columns (1/8 inch by 6 feet [0.32 by 183 cm]) filled with 3% OV-1 on Gas-Chrom Q (100 to 200 mesh) and coiled stainless-steel columns (1/8 inch by 6 feet [0.32 by 183 cm]) filled with GP 2% SP-1200/1% H3PO4 on Chromsorb WAW, Applied Science Laboratories; [14C]methanol (4 mCi/ mmol), New England Nuclear Corp.; carbodiimide [1ethyl-3-(3-dimethylaminopropyl)carbodiimide

Chemical characterization and immunogenicity of capsular polysaccharide isolated from mucoid Staphylococcus aureus

“…PCP from strain SAl mucoid was compared serologically with extracts from other well-characterized, encapsulated S. aureus strains. The capsular polysaccharide elaborated by strain SAl mucoid formed a precipitin line of identity with an extract from strain M (22,26) but not with the nonmucoid parent strain SAl or with two strains pro- ducing heterologous capsules, Smith diffuse (15) and NS58D (37) (Fig. 2).…”

“…The three S. aureus isolates whose capsules have been best characterized biologically and biochemically are strains M, T, and Smith diffuse (34). Strain M produces a capsule composed of 2-acetamido-2-deoxy-D-galacturonic acid, 2acetamido-2-deoxy-D-fucose, and taurine (22,26). Capsular * Corresponding author.…”

Transcriptional analysis of type 1 capsule genes in Staphylococcus aureus

“…Since its isolation, strain M has been used as a prototype of encapsulated staphylococci in a number of studies (for a review, see Wilkinson, 1983). The capsule of strain M is composed of taurine, 2-acetamido-2-deoxy-fucose, and 2-acetamido-2-deoxy-D-galacturonic acid in the molar ratios of 1:2:4 (Murthy et al, 1983;Liau and Hash, 1977). The presence of taurine in the capsule is dependent on the presence of taurine in the growth medium (Wilkinson et al, 1979).…”

Synthesis and immunologic properties in mice of vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides conjugated to Pseudomonas aeruginosa exotoxin A

“…These CPs have unusual properties which may account for the fact that this information emerged only recently. (i) S. aureus isolates from the blood of patients do not have the mucoid appearance usually associated with capsulation; widely studied capsulated S. aureus strains, especially the mucoid strain Smith, were not associated with bacteremia in humans (20,27,31,32,37,38,41,42,52,53). The amount of CP synthesized by bacteremic isolates is comparatively low; incubation of these strains with India ink does not demonstrate the CP as it does the large capsule surrounding the Smith strain (type 2 according to the classification of Kar- poor immunogens in humans, especially in patients with reduced resistance (15,47).…”