Characterization of the lipid A component of genuine smooth-form lipopolysaccharide

The smooth-form lipopolysaccharide of Salmonella abortus equi had earlier been separated into three distinct fractions, a long-chain fraction with an O chain containing 20-50 repeating units, a short-chain fraction consisting of an R lipopolysaccharide and another with 1-6 repeating units, and an R fraction identical to the lipopolysaccharide synthesized by Ra.b-mutant bacteria [Galanos et al. (1988) J. Chromatogr. 440, 397-404]. In this paper, the corresponding lipid A from each fraction was prepared by a newly elaborated procedure based on hydrolysis of the fractions in calcium acetate buffer (pH 3.5) followed by separation of the resulting free lipid A from the polysaccharide on a Sephadex G-100 column. Chemical analysis revealed that lipid A of the R fraction contained the expected spectrum and amounts of fatty acids and it proved to be structurally identical to lipid A of previously studied Salmonella R mutants. In contrast, the lipid A of the long-chain fraction contained only about 60% fatty acids compared to that of the R fraction. The lipid A of the short-chain fraction also expressed a reduced substitution pattern of acyl residues.