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Journal of Bacteriology, October 2007, p. 6945-6956, Vol. 189, No. 19
0021-9193/07/$08.00+0     doi:10.1128/JB.00500-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Flagellin Glycans from Two Pathovars of Pseudomonas syringae Contain Rhamnose in D and L Configurations in Different Ratios and Modified 4-Amino-4,6-Dideoxyglucose{triangledown}

Kasumi Takeuchi,1*,{dagger} Hiroshi Ono,2,{dagger} Mitsuru Yoshida,2 Tadashi Ishii,3 Etsuko Katoh,1 Fumiko Taguchi,4 Ryuji Miki,4 Katsuyoshi Murata,1 Hanae Kaku,5 and Yuki Ichinose4

National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602,1 National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642,2 Forestry and Forest Products Research Institute, 1, Matsunosato, Tsukuba, Ibaraki 305-8687,3 Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530,4 Faculty of Agriculture, Meiji University, Kawasaki, Kanagawa 214-8571, Japan5

Received 2 April 2007/ Accepted 10 July 2007

Flagellins from Pseudomonas syringae pv. glycinea race 4 and Pseudomonas syringae pv. tabaci 6605 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 nuclear magnetic resonance 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, ß-D-Quip4N(3-hydroxy-1-oxobutyl)2Me-(1->3)-{alpha}-L-Rhap-(1->2)-{alpha}-L-Rhap. Furthermore, mass analysis suggests that the glycans on each of the six serine residues are composed of similar trisaccharide units. Determination of the enantiomeric ratio of Rha from the flagellin proteins showed that flagellin from P. syringae pv. tabaci 6605 consisted solely of L-Rha, whereas P. syringae pv. glycinea race 4 flagellin contained both L-Rha and D-Rha at a molar ratio of about 4:1. Taking these findings together with those from our previous study, we conclude that these flagellin glycan structures may be important for the virulence and host specificity of P. syringae.

* Corresponding author. Mailing address: National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan. Phone: (81) 29 838 7005. Fax: (81) 29 838 7408. E-mail: kasumit{at}affrc.go.jp

{triangledown} Published ahead of print on 20 July 2007.

{dagger} K.T. and H.O. contributed equally to this work.

Journal of Bacteriology, October 2007, p. 6945-6956, Vol. 189, No. 19
0021-9193/07/$08.00+0     doi:10.1128/JB.00500-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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