Structural studies on the linkage unit between poly(N-acetylglucosamine 1-phosphate) and peptidoglycan in cell walls of Bacillus pumilus AHU 1650

Abstract

Structural studies were carried out on the polymer chains and their linkage regions in two kinds of teichoic acids, poly(N-acetylglucosamine 1-phosphate) [poly(GlcNAc-1-P)] and glycerol teichoic acid, bound to peptidoglycan in the cell walls of Bacillus pumilus AHU 1650. The poly(GlcNAc-1-P)-glycan complex isolated from lysozyme digests of the cell walls contained mannosamine and glycerol as minor components. On the basis of proton NMR spectroscopic data and isolation of N-acetylglucosamine 4-phosphate from acid hydrolysates, the poly(GlcNAc-1-P) was shown to be a polymer in which N-acetylglucosamine 1-phosphate units are joined at C-4 of the glucosamine residues. Mild alkaline hydrolysis of the poly(GlcNAc-1-P)-glycan complex gave a mannosamine-linked glycan fragment and the acidic polymer fraction that contained glycerol residues. Mild acid treatment of the mannosamine-linked glycan fragment gave the linkage disaccharide, ManNAc(beta 1----4)GlcNAc, whereas the acidic polymer fraction was degraded by this treatment into N-acetylglucosamine 4-phosphate and a glycerol-containing fragment characterized as P-(Gro-P)7 (Gro = glycerol). On the other hand, direct mild acid hydrolysis of the complex gave a fragment characterized as P-(Gro-P)7-ManNAc(beta 1----4)GlcNAc. These results lead to a conclusion that in the cell walls the poly(GlcNAc-1-P) chain is attached to peptidoglycan through a linkage unit, (Gro-P)7-ManNAc(beta 1----4)GlcNAc. By means of similar procedures, it was shown that the other cell wall polymer, glycerol teichoic acid, is also attached to peptidoglycan through the same disaccharide, ManNAc(beta 1----4)GlcNAc.