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Glycobiology Advance Access published online on January 13, 2009
Glycobiology, doi:10.1093/glycob/cwn155
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Published by Oxford University Press 2009.

Communications

Structural Elucidation of the Novel Core Oligosaccharide from LPS of Burkholderia cepacia Serogroup O4

Hussein Masoud1,2, Malcolm B. Perry1, Jean-Robert Brisson1, Dusan Uhrin1,3, Jianjun Li1 and James C. Richards1,*

1 Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, K1A 0R6, Canada
2 Department of Biological Sciences, Faculty of Science, University of Jordan, 11942 Amman, Jordan
3 Department of Chemistry, The University of Edinburgh, Edinburgh, EH9 3JJ, UK

* To whom correspondence should be addressed at the Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada, K1A 0R6. Tel (613) 993–7506 Fax (613) 941–1327, E-mail: Jim.Richards{at}nrc-cnrc.gc.ca

Lipopolysaccharide (LPS) is an important virulence factor of Burkholderia cepacia, an opportunistic bacterial pathogen causing life-threatening disease in cystic fibrosis patients and the immunocompromised. B. cepacia LPS is comprised of an O-specific polysaccharide covalently linked to a core oligosaccharide (OS) which in turn is attached to a lipid A moiety. The complete structure of the LPS core oligosaccharide from B. cepacia serotype O4 was investigated by detailed NMR and mass spectrometry (MS) methods. High (HMW) and low molecular weight (LMW) OSs were obtained by deacylation, dephophorylation and reducing-end reduction of the LPS. Glycan and NMR analyses established that both OSs contain a common inner-core structure comprised of D-glucose, L-glycero-D-manno-heptose, D-glycero-D-manno-heptose, 3-deoxy-D-manno-octulsonic acid and D-glycero-D-talo-2-octulosonic acid. The structure of the LMW OS differed from that of the HMW OS in that it lacks a tetra-rhamnosyl GlcNAc OS extension. These structural conclusions were confirmed by tandem MS analyses of the two OS fractions as well as an OS fraction obtained by alkaline deacylation of the LPS. The location of a phosphoethanolamine substituent in the core region was determined by ESI-MS and methylation analysis of O-deacylated LPS and core OS samples. Polyclonal antibody to B. cepacia serotype O4 core OS was cross-reactive with several other serotypes indicating common structural features.

Key words: Burkholderia cepacia / Lipopolysaccharide / Core region / D-glycero-D-talo-2-octulosonic acid


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