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J. Biol. Chem., Vol. 279, Issue 35, 36470-36480, August 27, 2004

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Investigation of the Structural Requirements in the Lipopolysaccharide Core Acceptor for Ligation of O Antigens in the Genus Salmonella

WAAL "LIGASE" IS NOT THE SOLE DETERMINANT OF ACCEPTOR SPECIFICITY^*

Natalia A. Kaniuk, Evgeny Vinogradov¶, and Chris Whitfield||

From the Department of Microbiology, University of Guelph, Guelph, Ontario N1G 2W1 and ^¶Institute for Biological Sciences, National Research Council, Ottawa, Ontario K1A 0R6, Canada

The ligation of O antigen polysaccharide to lipid A-core oligosaccharide is a late step in the formation of the complex glycolipid known as lipopolysaccharide. Although the process has been localized to the periplasmic face of the inner membrane, details of the ligation mechanism have not been resolved. To date, there is only one gene product (WaaL, often referred to as "ligase") known to be required. There exists a requirement for a specific lipid A-core oligosaccharide acceptor structure for ligation activity, and it has been proposed that the WaaL protein imparts this acceptor specificity. Here the structural requirements in the core oligosaccharide acceptor for O antigen ligation are investigated in prototype serovars of Salmonella enterica. Complementation experiments in mutants with defined core oligosaccharide structure indicate that the specificity of the ligation reaction for a particular core oligosaccharide structure is not dependent on the WaaL protein alone. The data provide the first indication of a more complicated recognition process involving additional cellular components.

Received for publication, February 6, 2004 , and in revised form, June 2, 2004.

^* This work was supported in part through funding by the Canadian Bacterial Disease Network (to C. W.) and the National Sciences and Engineering Research Council. 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.

Recipient of a National Sciences and Engineering Research Council PGS-B graduate scholarship.

^|| To whom correspondence should be addressed. Tel.: 519-824-4120 (ext. 53361); Fax: 519-837-1802; E-mail: cwhitfie{at}uoguelph.ca.

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