Size heterogeneity of Salmonella typhimurium lipopolysaccharides in outer membranes and culture supernatant membrane fragments

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Abstract
Enterobacteriaceae cells growing in liquid media shed fragments of their outer membranes. These fragments, which may constitute a biologically important form of gram-negative bacterial endotoxin, were reported to contain proteins, phospholipids and lipopolysaccharides (LPS). In this study the sizes of LPS molecules in shed membrane fragments and outer membranes from cells growing in broth cultures were compared. Using conditional mutants of S. typhimurium which incorporate specific sugars into LPS, radiolabeled LPS was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This technique revealed that S. typhimurium LPS are more heterogeneous than previously known; molecules possessing from 0 to more than 30 O-chain repeat units were identified in outer membranes, supernatant fragments and purified LPS. The size distributions of LPS molecules in outer membranes and supernatant fragments were similar; supernatant fragments appeared to be slightly enriched in molecules with long O-polysaccharide chains. Apparently, LPS molecules of many sizes are synthesized, translocated to outer membranes and released into culture supernatants. Since the hydrophilic O-polysaccharides extend from bacterial surfaces into the aqueous environment, the cell surface topography of this bacterium may be very irregular. The heterogeneity in the degree of polymerization of O-antigenic side chains may influence the interactions of the toxic moiety of LPS (lipid A) with host constituents.

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