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Glycobiology, 2001, Vol. 11, No. 12 1009-1015
© 2001 Oxford University Press

Glycine is a common substituent of the inner core in Haemophilus influenzae lipopolysaccharide

Jianjun Li2, Sebastian H.J. Bauer3, Martin Månsson3, E. Richard Moxon4, James C. Richards2 and Elke K.H. Schweda1,3

2Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada K1A OR6; 3Clinical Research Centre, Karolinska Institutet and University College of South Stockholm, NOVUM, S-141 86 Huddinge, Sweden; 4Molecular Infectious Diseases Group, University of Oxford Department of Pediatrics, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford 0X3 9DS, UK

A survey of both typeable and nontypeable strains of Haemophilus influenzae indicated that they contain glycine (Gly) in their lipopolysaccharide (LPS). Significant amounts (30–250 pmol Gly/µg LPS) were determined by high-performance anion-exchange chromatography using pulsed amperometric detection after treatment of the LPS with mild alkali. Oligosaccharides obtained from LPS after mild acid hydrolysis and gel filtration chromatography were investigated by electrospray ionization mass spectrometry (ESI-MS) and capillary electrophoresis (CE) ESI-MS. In all cases, molecular ions corresponding to the major glycoforms were identified and were accompanied by ions differing by 57 Da, thus indicating the presence of glycine. The position of glycine in these glycoforms was determined by CE-ESI-MS/MS analyses. It was found that, depending on strain, glycine can substitute each of the heptoses of the inner-core element, L-{alpha}-D-Hepp-(1->2)-[PEtn->6]-L-{alpha}-D-Hepp-(1->3)-L-{alpha}-D-Hepp-(1->5)-{alpha}-Kdo of H. influenzae LPS as well as Kdo. In some strains, mixtures of monosubstituted Gly-containing glycoforms having different substitution patterns were identified.

1 To whom correspondence should be addressed


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