ProGlyProt ID BC172 Organism Information Organism Name Pseudomonas 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 Gene Information Gene Name fliC NCBI Gene ID Protein Information Protein Name Flagellin UniProtKB/SwissProt ID Q6I809 NCBI RefSeq EMBL-CDS BAD24670.1 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 Location Surface 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. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Ser) linked Experimentally Validated Glycosite(s) in Full Length Protein S143, S164, S176, S183, S193, S201 Experimentally Validated Glycosite(s ) in Mature Protein S143, 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 seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Rapid 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 tri 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 [H Protein Glycosylation linked (PGL) gene(s) OST Gene Name OST NCBI Gene ID OST GenBank Gene Sequence OST Protein Name OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) OST Genome Context Characterized Accessory Gene(s) FGT1 and FGT2 are two glycosyltransferases which are encoded in the flagellin glycosylation island. Non-glycosylated and partially glycosylated flagellins were respectively produced by fgt1 and fgt2 mutants. PGL Additional Links CAZy Literatures Reference(s) 1) 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] 2) 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 syringa Additional Comments Year of Identification 2003 Year of Validation 2006