Abstract
The cell walls of Staphylococcus aureus H were found to contain mannosamine in an amount (28.1 nmol/mg) comparable to the content of muramic acid 6-phosphate. The acidic polymer fraction obtained by heating the cell walls at pH 2.5 was shown to contain mannosamine and glycerol in addition to the components of ribitol teichoic acid. Mild alkali treatment of this polymer fraction followed by gel filtration resulted in separation of a disaccharide N-acetylmannosaminyl(1 leads to 4)N-acetylglucosamine and the ribitol teichoic acid moiety that contained glycerol. Smith degradation of the reduction product from the same polymer fraction gave a fragment characterized as (1,2-ethylene-diol phosphate)-(glycerol phosphate)3-N-acetylmannosaminyl(1 leads to 4)N-acetylxylosaminitol. Thus, the ribitol teichoic acid chain in the cell walls is probably linked to peptidoglycan through a linkage unit, (glycerol phosphate)3-N-acetylmannosaminyl(1 leads to 4)N-acetylglucosamine.
Highlights
Smith Degradation-The TA-S preparation (20 pmol of phosphooccur in the cell walls from several Bacillus species as the rus) after reduction with N&H4 was oxidized with 0.1 M NaI04 in commonlinkage units betweenpeptidoglycan and glycerol 0.5 ml of 0.1 M sodium acetate buffer, pH 5.0, in the dark at 4 "C for teichoic acids whicahre different in the structure of glycosidic branches and backbone chains? In addition, another type of linkage disaccharide, GlcS(1 + 4)GlcNAc, was indicated to be present in the glycerol teichoic acid-glycopeptide complex
6-phosphate when the cell walls werexamined without heating at pH 2.5. This result suggests that the heating of cell walls at pH 2.5 led to selectivecleavageof the linkagebetween the teichoicacid moiety and thephosphate group of a muramic acid 6-phosphate residue in the peptidoglycan moiety
The above acetylglucosaminerecovered in Peak 3 was decreased to one- result indicates that the mannosamine residue in this fragthird. These results strongly suggest that thelinkage unit of ment was substituted at C-3 or C-4 probably by a phosphoryl the monosaccharide-type was absent from the cell wallsof S. group
Summary
Smith Degradation-The TA-S preparation (20 pmol of phosphooccur in the cell walls from several Bacillus species as the rus) after reduction with N&H4 was oxidized with 0.1 M NaI04 in commonlinkage units betweenpeptidoglycan and glycerol 0.5 ml of 0.1 M sodium acetate buffer, pH 5.0, in the dark at 4 "C for teichoic acids whicahre different in the structure of glycosidic branches and backbone chains? The abbreviations used are: GlcNAc, N-acetylglucosamine; ManNAc, N-acetylmannosamine; P, phosphate group; TA-S, teichoic acid-linked saccharide. This result suggests that the heating of cell walls at pH 2.5 led to selectivecleavageof the linkagebetween the teichoicacid moiety and thephosphate group of a muramic acid 6-phosphate residue in the peptidoglycan moiety.
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