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Glycobiology Advance Access published online on November 10, 2004
Glycobiology, doi:10.1093/glycob/cwi015
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Received July 13, 2004
Revised October 1, 2004
Accepted November 5, 2004

ORIGINAL ARTICLES

Structural analysis of the lipopolysaccharide from Pasteurella multocida genome strain Pm70 and identification of the putative lipopolysaccharide glycosyltransferases

Frank St. Michael 1, Evgeny Vinogradov 1, Jianjun Li 1, and Andrew D. Cox 1*

1 Institute for Biological Sciences, National Research Council, Ottawa, ON, Canada, K1A 0R6

* To whom correspondence should be addressed.
Andrew D. Cox, E-mail: Andrew.Cox{at}nrc-cnrc.gc.ca


  Abstract

Pasteurella multocida is an important multi-species veterinary pathogen. The cell surface lipopolysaccharide (LPS) is an important virulence factor and forms the basis of the serotyping scheme, although little structural information about the LPS is known. The structure of the LPS from the Pasteurella multocida genome strain Pm70 was elucidated in this study. The LPS was subjected to a variety of degradative procedures. The structures of the purified products were established by monosaccharide and methylation analyses, NMR spectroscopy and mass spectrometry. The following structure for the core oligosaccharide was determined on the basis of the combined data from these experiments, where based on the NMR data all sugars were found in pyranose ring forms. Glucose, galactose and N-acetyl-galactosamine residues were all present as D-isomers. Kdo is 2-keto-3-deoxy-octulosonic acid, L-{alpha}-D-Hep is L-glycero-D-manno-heptose, and PEtn is phosphoethanolamine. Identification of the core oligosaccharide structure enabled a search for glycosyltransferase homologues in the Pm70 genome, and revealed a clustering of the genes putatively responsible for outer core oligosaccharide biosynthesis.

Keywords: Pasteurella multocida; LPS; Core oligosaccharide; Mass spectrometry; NMR.
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