O-acetylation affects the binding properties of the carboxyl groups on the Vi bacterial polysaccharide

The capsular polysaccharide of Salmonella typhi and Citrobacter freundii (Vi) is a linear homopolymer of (1-->4)-linked 2-acetamido-2-deoxy-alpha-D-galactopyranosyluronic acid partially O-acetylated at the C-3 position. The physico-chemical properties of the carboxyl groups of the Vi polysaccharide, as a function of different degrees of O-acetylation, were studied by potentiometric titration, circular dichroism, and their reaction with the bulky nucleophile 2-nitro-phenylhydrazine (NPH). Potentiometric titrations with K+ and Ca2+ hydroxides showed that the difference in the free energy of binding between the two cations (delta GKCa) was inversely proportional to the degree of O-acetylation. Similar cationic effects were found when measuring circular dichroism. Moreover there was also an inverse relation between the degree of O-acetylation and the extent of binding of NPH to the carboxyl groups. These data all indicate that O-acetyl groups hinder the association of carboxyls with cations and nucleophilic reagents. This provides a possible explanation for the importance of the O-acetyl and the relative unimportance of the carboxyl groups in contributing to the immunologic properties of the Vi.