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J. Bacteriol. doi:10.1128/JB.00500-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Title: Flagellin glycans from two pathovars of Pseudomonas syringae contain rhamnose in D- and L- configurations at different ratios and modified 4-amino-4,6-dideoxyglucose

National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.; National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.; Forestry and Forest Products Research Institute, 1, Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.; Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530 Japan.; Faculty of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan

^* To whom correspondence should be addressed. Email: kasumit{at}affrc.go.jp.

	Abstract

Flagellins from Pseudomonas syringae pv. glycinea race 4 (Pgl4) and P. s. pv. tabaci 6605 (Pta6605) have been found to be glycosylated. Glycosylation of flagellin is essential for bacterial virulence and is also involved in the determination of host specificity. Flagellin glycans from both pathovars were characterized and common sites of glycosylation were identified on six serine residues (positions 143, 164, 176, 183, 193 and 201). The structure of the glycan at serine 201 (S201) of flagellin from each pathovar was determined by sugar composition analysis, mass spectrometry and ¹H and ¹³C NMR spectroscopy. These analyses showed that the S201 glycans from both pathovars were composed of a common unique trisaccharide consisting of two rhamnosyl (Rha) residues and one modified 4-amino-4,6-dideoxyglucosyl (Qui4N) residue, -D-Quip4N(3-hydroxy-1-oxobutyl)2Me-(13)--L-Rhap-(12)--L-Rhap. Furthermore, mass analysis suggests that the glycans on each of the six serine residues are composed of a similar trisaccharide unit. Determination of the enantiomeric ratio of Rha from the flagellin proteins showed that flagellin from Pta6605 consisted solely of L-Rha, whereas Pgl4 flagellin contained both L-Rha and D-Rha at a molar ratio of about 4:1. Together with our previous study, we conclude that these structures of the flagellin glycans may be important for virulence and host specificity of P. syringae.

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