Some immunochemical and chemical aspects of S and R Shigella dysenteriae 1 endotoxins.

Endotoxins of S and R forms of Shigella dysenteriae 1 were prepared by NaCl-Na citrate extraction, purified by gel chromatography on Sephadex G 200 and on Sepharose 4B and subjected to immunochemical and chemical analysis. The toxins contained 25--30% of lipids, 40--50% of carbohydrates and 14--24% of protein. The lipid and protein moieties of the lipopolysaccharide-protein complexes exhibited no significant difference, whereas the sugar moieties differed markedly (both qualitatively and quantitatively), in relation to the growth form of the culture. The lipid moiety, which consists at least of 22 fatty acids, has the greatest relative content (approx. 50%) of behenic acid, 22:0, and palmitic acid, 16:0 (approx. 11%). In the protein moiety, at least 16 amino acids were determined; these amino acids were identical in both endotoxin types, but their total content was higher in the R form, giving an R:S ratio of 1.7 +/- 0.2. The sugar moiety consists of galactose, glucosamine and either rhamnose (in S endotoxin) or aldoheptose (in R endotoxin). The difference of the chemical composition of the sugar moiety is believed to account for the diametric difference in the immunochemical character, in particular the different behaviour in the electric field, of both endotoxin types. The average content of 3-deoxy-D-manno-2-octulosonic acid was determined as 0.5% for both S and R endotoxin. Trace amounts of O-phosphorylethanolamine were found. Individual aspects of the chemical and immunochemical analysis are discussed in detail.