Bordetella bronchiseptica is a pathogen of humans and animals that colonizes the respiratory tract. It produces a lipopolysaccharide O antigen that contains a homopolymer of 2,3-dideoxy-2,3-diacetamido-l-galacturonic acid (l-GalNAc3NAcA). Some of these sugars are found in the uronamide form (l-GalNAc3NAcAN), and there is no discernible pattern in the distribution of amides along the chain. A B. bronchiseptica wbmE mutant expresses an O polysaccharide unusually rich in uronamides. The WbmE protein localizes to the periplasm and catalyzes the deamidation of uronamide-rich O chains in lipopolysaccharide purified from the mutant, to attain a wild-type uronamide/uronic acid ratio. WbmE is a member of the papain-like transglutaminase superfamily, and this categorization is consistent with a deamidase role. The periplasmic location of WbmE and its acceptance of complete lipopolysaccharide as substrate indicate that it operates at a late stage in lipopolysaccharide biosynthesis, after polymerization and export of the O chain from the cytoplasm. This is the first report of such a modification of O antigen after assembly. The expression of wbmE is controlled by the Bordetella virulence gene two-component regulatory system, BvgAS, suggesting that this deamidation is a novel mechanism by which these bacteria modify their cell surface charge in response to environmental stimuli.
lipopolysaccharide biosynthesis, O polysaccharide, modification, Bordetella bronchiseptica, periplasm, WbmE
NCBI PubMed ID: 19015265Publication DOI: 10.1074/jbc.M807729200Journal NLM ID: 2985121RPublisher: Baltimore, MD: American Society for Biochemistry and Molecular Biology
Correspondence: jeguel@gmail.com
Institutions: Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, United Kingdom, Department of Veterinary Medicine, University of Cambridge, United Kingdom
Methods: SDS-PAGE, DNA techniques, genetic methods, CE-MS
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