ProGP227

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ProGP ID ProGP227
Validation Status Characterized
Organism Information
Organism NamePseudomonas syringae pv. tabaci 6605
Domain Bacteria
Classification Family: Pseudomonadaceae
Order: Pseudomonadales
Class: Gammaproteobacteria
Division or phylum: "Proteobacteria"
Taxonomic ID (NCBI) 322
Genome Sequence(s)
EMBL AB061230
Organism Additional Information This Gram-negative bacterium is phytopathogenic causing wildfire disease in tobacco plants. In this host tobacco infection, flagella play an important role.
Gene Information
Gene NamefliC
Protein Information
Protein NameFlagellin
UniProtKB/SwissProt ID Q6I809
EMBL-CDSBAD24670.1
UniProtKB Sequence >tr|Q6I809|Q6I809_PSESZ Flagellin OS=Pseudomonas syringae pv. tabaci GN=fliC PE=4 SV=1 MALTVNTNVASLNVQKNLGRASDALSTSMTRLSSGLKINSAKDDAAGLQIATKITSQIRG QTMAIKNANDGMSLAQTAEGALQESTNILQRMRELAVQSRNDSNSSTDRDALNKEFTAMS SELTRIAQSTNLNGKNLLDGSASTMTFQVGSNSGASNQITLTLSASFDANTLGVGSAVTI AGSDSTTAETNFSAAIAAIDSALQTINSTRADLGAAQNRLTSTISNLQNINENASAALGR VQDTDFAAETAQLTKQQTLQQASTSVLAQANQLPSAVLKLLQ
Sequence length 282 AA
Subcellular LocationSurface
Function Flagellin, the major component of the flagella filament functions as one of the pathogen-associated molecular patterns (PAMPs). It is a major HR (hypersensitive reaction) elicitor in this pathogen but does not remarkably induce hypersensitive cell death in its host tobacco plants.
Glycosylation Status
Glycosylation Type O (Ser) linked
Experimentally Validated Glycosite(s) in Full Length ProteinS143, S164, S176, S183, S193, S201
Experimentally Validated Glycosite(s ) in Mature ProteinS143, S164, S176, S183, S193, S201
Glycosite(s) Annotated Protein Sequence >tr|Q6I809|Q6I809_PSESZ Flagellin OS=Pseudomonas syringae pv. tabaci GN=fliC PE=4 SV=1 MALTVNTNVASLNVQKNLGRASDALSTSMTRLSSGLKINSAKDDAAGLQIATKITSQIRG QTMAIKNANDGMSLAQTAEGALQESTNILQRMRELAVQSRNDSNSSTDRDALNKEFTAMS SELTRIAQSTNLNGKNLLDGSAS*(143)TMTFQVGSNSGASNQITLTLS*(164)ASFDANTLGVGS*(176) AVTI AGS*(183)DSTTAETNFS*(193)AAIAAIDS*(201)ALQTINSTRADLGAAQNRLTSTISNLQNINENASAALGR VQDTDFAAETAQLTKQQTLQQASTSVLAQANQLPSAVLKLLQ
Sequence Around Glycosites (21 AA) NGKNLLDGSASTMTFQVGSNS
GASNQITLTLSASFDANTLGV
SFDANTLGVGSAVTIAGSDST
GVGSAVTIAGSDSTTAETNFS
SDSTTAETNFSAAIAAIDSAL
NFSAAIAAIDSALQTINSTRA
Glycosite Sequence Logo
Technique(s) used for Glycosylation DetectionRapid migration on SDS-PAGE after chemical deglycosylation with TFMS, glycoprotein staining using GelCode® glycoprotein staining kit.
Technique(s) used for Glycosylated Residue(s) Detection Site-directed mutagenesis and MALDI-TOF MS (matrix assisted laser desorption/ionization time of flight mass spectrometry) analysis
Protein Glycosylation- Implication Glycosylation, especially at positions S176 and S183, is required for smooth motility and bacterial virulence. Glycosylation is one of the important process in bacterial HR-inducing (hypersensitive reaction, a rapid and strong plant defence response) activity. Glycosylation may also serve to mask elicitor function of flagellin molecule in order to avoid plant recognition. Glycosylation of sites that are most surface-exposed plays a role in the swarming motility of the P. syringae pv. tabaci 6605.
Glycan Information
Glycan Annotation Linkage: α-L-Rha-Ser.
Tetrasaccharide, 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methyl-Glcp-(1→3)-α-L-Rhap-(1→2)-α-L-Rhap-(1→2)-α-L-Rha-(1→, as well as a trisaccharide, 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methyl-Glcp-(1→3)-α-L-Rhap-(1→2)-α-L-Rha-(1→. Rhamnose (Rha) is exclusively of the L form. 4,6-dideoxy-4-(3-hydroxy-3-methylbutanamido)-2-O-methylglucopyranose is named as viosamine. Therefore, in the glycans, modified viosamine is present.
The ratios of a trisaccharide and a tetrasaccharide on LC by fluorescence detection were about 5:1, indicating that about five serine residues (including S201) were glycosylated with a trisaccharide. The major component of each glycan is similar to that at S201.
Microheterogeneity in glycosylation is observed.
BCSDB ID24060
Technique(s) used for Glycan Identification Sugar composition analysis using an ABEE (p-aminobenzoic acid ethyl ester) labeling kit, MALDI-TOF MS (matrix-assisted laser desorption ionization–time-of-flight mass spectrometry), determination of D-Rha/L-Rha ratios using gas chromatography (GC), and 1H-NMR analyses including 1H-1H correlation spectra [DQF-COSY (double quantum filtered correlation spectroscopy), TOCSY (total correlation spectroscopy), and NOESY (nuclear Overhauser and exchange spectroscopy)] and 1H-13C correlation spectra [HSQC (heteronuclear single-quantum coherence) and HMBC (heteronuclear multiple bond connectivity)].
Literature
Year of Identification2003
Year of Identification Month Wise2003.3.1
Year of Validation 2006
Reference Konishi, T., Taguchi, F., Iwaki, M., Ohnishi-Kameyama, M., Yamamoto, M., Maeda, I., Nishida, Y., Ichinose, Y., Yoshida, M. and Ishii, T. (2009) Structural characterization of an O-linked tetrasaccharide from Pseudomonas syringae pv. tabaci flagellin. Carbohydr Res, 344, 2250-2254. [PubMed: 19766989]
Author Konishi, T., Taguchi, F., Iwaki, M., Ohnishi-Kameyama, M., Yamamoto, M., Maeda, I., Nishida, Y., Ichinose, Y., Yoshida, M. Ishii, T.
Research GroupUnit for Laboratory Animal Medicine and Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Corresponding Author Ishii, T.
ContactUnit for Laboratory Animal Medicine and Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Reference Takeuchi, K., Ono, H., Yoshida, M., Ishii, T., Katoh, E., Taguchi, F., Miki, R., Murata, K., Kaku, H. and Ichinose, Y. (2007) 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. J Bacteriol, 189, 6945-6956. [PubMed: 17644592]
Author Ichinose, Y.
Research GroupNational Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan. kasumit@affrc.go.jp
Corresponding Author Takeuchi, K., Ono, H., Yoshida, M., Ishii, T., Katoh, E., Taguchi, F., Miki, R., Murata, K., Kaku, H. Ichinose, Y.
ContactNational Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan. kasumit@affrc.go.jp
Reference Taguchi, F., Takeuchi, K., Katoh, E., Murata, K., Suzuki, T., Marutani, M., Kawasaki, T., Eguchi, M., Katoh, S., Kaku, H. et al. (2006) Identification of glycosylation genes and glycosylated amino acids of flagellin in Pseudomonas syringae pv. tabaci. Cell Microbiol, 8, 923-938. [PubMed: 16681835]
Author Taguchi, F., Takeuchi, K., Katoh, E., Murata, K., Suzuki, T., Marutani, M., Kawasaki, T., Eguchi, M., Katoh, S., Kaku, H. et al
Research GroupThe Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan.
Corresponding Author Yuki Ichinose
ContactThe Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan.
Reference Konishi, T., Taguchi, F., Iwaki, M., Ohnishi-Kameyama, M., Yamamoto, M., Maeda, I., Nishida, Y., Ichinose, Y., Yoshida, M. and Ishii, T. (2009) Structural characterization of an O-linked tetrasaccharide from Pseudomonas syringae pv. tabaci flagellin. Carbohydr Res, 344, 2250-2254. [PubMed: 19766989]
Author Konishi, T., Taguchi, F., Iwaki, M., Ohnishi-Kameyama, M., Yamamoto, M., Maeda, I., Nishida, Y., Ichinose, Y., Yoshida, M. Ishii, T.
Research GroupForestry and Forest Products Research Institute, 1, Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.
Corresponding Author Ishii, T.
ContactForestry and Forest Products Research Institute, 1, Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.
Reference1) Taguchi, F., Yamamoto, M., Ohnishi-Kameyama, M., Iwaki, M., Yoshida, M., Ishii, T., Konishi, T. and Ichinose, Y. (2010) Defects in flagellin glycosylation affect the virulence of Pseudomonas syringae pv. tabaci 6605. Microbiology, 156, 72-80. [PubMed: 19815579]
AuthorTaguchi, F., Yamamoto, M., Ohnishi-Kameyama, M., Iwaki, M., Yoshida, M., Ishii, T., Konishi, T. and Ichinose, Y.
Research GroupGraduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530 Japan.
Corresponding Author Ichinose, Y
ContactGraduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530 Japan.
Reference Taguchi, F., Shimizu, R., Inagaki, Y., Toyoda, K., Shiraishi, T. and Ichinose, Y. (2003) Post-translational modification of flagellin determines the specificity of HR induction. Plant Cell Physiol, 44, 342-349. [PubMed: 12668781]
Author Taguchi, F., Shimizu, R., Inagaki, Y., Toyoda, K., Shiraishi, T. Ichinose, Y.
Research GroupLaboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University, Tsushima-naka 1-1-1, Okayama, 700-8530 Japan
Corresponding Author Ichinose, Y.
ContactLaboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University, Tsushima-naka 1-1-1, Okayama, 700-8530 Japan