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J. Biol. Chem., Vol. 281, Issue 26, 18135-18144, June 30, 2006

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Complete Structures of Bordetella bronchiseptica and Bordetella parapertussis Lipopolysaccharides^*

Andrew Preston, Bent O. Petersen, Jens Ø. Duus, Joanna Kubler-Kielb^¶, Gil Ben-Menachem^¶, Jianjun Li^||, and Evgeny Vinogradov^||1

From the ^||Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A OR6, Canada, Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada, Carlsberg Laboratory, DK 2500 Valby Copenhagen, Denmark, and ^¶NICHD, National Institutes of Health, Bethesda, Maryland 20892

The structures of the lipopolysaccharide (LPS) core and O antigen of Bordetella bronchiseptica and Bordetella parapertussis are known, but how these two regions are linked to each other had not been determined. We have studied LPS from several strains of these microorganisms to determine the complete carbohydrate structure of the LPS. LPS was analyzed using different chemical degradations, NMR spectroscopy, and mass spectrometry. This identified a novel pentasaccharide fragment that links the O chain to the core in all the LPS studied. In addition, although the O chain of these bacteria was reported as a homopolymer of 1,4-linked 2,3-diacetamido-2,3-dideoxy--galacturonic acid, we discovered that the polymer contains several amidated uronic acids, the number of which varies between strains. These new data describe the complete structure of the LPS carbohydrate backbone for both Bordetella species and help to explain the complex genetics of LPS biosynthesis in these bacteria.

Received for publication, December 30, 2005 , and in revised form, April 21, 2006.

^* Work in the Preston laboratory is supported in part by a Natural Sciences and Engineering Research Council Discovery Grant. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Inst. for Biological Sciences, National Research Council, 100 Sussex Dr., Ottawa, Ontario K1A 0R6, Canada. Tel.: 613-990-0832; Fax: 613-952-9092; E-mail: evguenii.vinogradov{at}nrc-cnrc.gc.ca.

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