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A more recent version of this article appeared on November 1, 2007

Papers In Press, published online ahead of print August 16, 2007
J. Lipid Res., doi:10.1194/jlr.M700193-JLR200
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Submitted on April 23, 2007
Revised on August 14, 2007
Accepted on August 16, 2007

A rapid, small-scale procedure for the structural characterization of lipid A applied to Citrobacter and Bordetella strains: discovery of a new structural element

Alina Tirsoaga, Asmaa El hamidi, Malcolm B. Perry, Martine Caroff, and Alexey Novikov

IBBMC, Orsay 91405

Corresponding Author: alexey.novikov{at}u-psud.fr

Endotoxins (lipopolysaccharides, LPSs) are part of the outer cell membrane of Gram-negative bacteria; their biological activities are associated mainly with the lipid component (lipid A) and even more specifically with discrete aspects of their fine structure. The need for a rapid and small-scale analysis of lipid A motivated us to develop a procedure that combines direct isolation of lipids A from bacterial cells with sequential release of their ester-linked fatty acids by a mild alkali treatment followed by MALDI-MS analysis. The method avoids the multi-step LPS extraction procedure and lipid A isolation. The whole process can be performed in a working day and applied to lyophilized bacterial samples as small as 1 milligram. We illustrate the method by applying it to the analysis of lipids A of three species of Citrobacter which were found to be identical. On the other hand, when applied to two batches of Bordetella bronchiseptica strain 4650, it highlighted the presence in one of them, of hitherto unreported hexosamine residues substituting the lipid A phosphate groups, possibly a new camouflage opportunity to escape a host defense system.


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 J. Bacteriol.Home page
N. Marr, A. Tirsoaga, D. Blanot, R. Fernandez, and M. Caroff
Glucosamine Found as a Substituent of Both Phosphate Groups in Bordetella Lipid A Backbones: Role of a BvgAS-Activated ArnT Ortholog
J. Bacteriol., June 15, 2008; 190(12): 4281 - 4290.
[Abstract] [Full Text] [PDF]


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