Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1388000 | Carbohydrate Research | 2013 | 7 Pages |
•We followed the O-antigen composition of S. Typhimurium by MAS NMR of intact cells.•The O-antigen of S. Typhimurium is partially de-O-acetylated in the stationary growth phase.•The de-O-acetylated O-antigen is due to hydrolysis of the O-acetylated O-antigen.•The de-O-acetylation is caused by the metabolism-induced basic pH of the medium.•The labile O-acetylation of the O-antigen generates non-stoichiometric O-acetylation states.
NMR spectroscopy can detect biomolecules like lipopolysaccharide directly on the surface of the cell, thus avoiding isolation and purification, and providing a more realistic description than the one derived from in vitro studies. Here we present a high-resolution magic-angle spinning NMR study of the O-antigen of Salmonella enterica serovar Typhimurium (S. Typhimurium) performed directly on the cells showing the alteration of its acetylation state over time. The O-antigen region of S. Typhimurium consists of the repeating unit [→2)-α-d-Manp-(1→4)-α-l-Rhap-(1→3)-α-d-Galp-(1→] where Man stands for mannose, Rha for rhamnose, and Gal for galactose. Man is substituted with abequose (Abe) O-acetylated at carbon 2. Our studies revealed that the appearance of de-O-acetylated O-antigen in the stationary growth phase is due to the de-O-acetylation of already synthesized O-acetylated O-antigen and that this reaction is caused by the metabolism-induced basic pH of the growth medium. The labile O-acetylation of the O-antigen we observed in S. Typhimurium generates non-stoichiometric O-acetylation states and therefore changes the nature of an immunogenic epitope.
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