ProGlyProt ID BC168 Organism Information Organism Name Pseudomonas aeruginosa PAK /5939 Domain Bacteria Classification Family: Pseudomonadaceae Order: Pseudomonadales Class: Gammaproteobacteria Division or phylum: "Proteobacteria" Taxonomic ID (NCBI) 287 Genome Sequence (s) EMBL M57501 Gene Information Gene Name fliC NCBI Gene ID Protein Information Protein Name Type a Flagellin UniProtKB/SwissProt ID P21184 NCBI RefSeq EMBL-CDS AAA63458.1 UniProtKB Sequence >sp|P21184|FLICA_PSEAE A-type flagellin OS=Pseudomonas aeruginosa GN=fliC PE=1 SV=3 MALTVNTNIASLNTQRNLNNSSASLNTSLQRLSTGSRINSAKDDAAGLQIANRLTSQVNG LNVATKNANDGISLAQTAEGALQQSTNILQRMRDLSLQSANGSNSDSERTALNGEVKQLQ KELDRISNTTTFGGRKLLDGSFGVASFQVGSAANEIISVGIDEMSAESLNGTYFKADGGG AVTAATASGTVDIAIGITGGSAVNVKVDMKGNETAEQAAAKIAAAVNDANVGIGAFSDGD TISYVSKAGKDGSGAITSAVSGVVIADTGSTGVGTAAGVTPSATAFAKTNDTVAKIDIST AKGAQSAVLVIDEAIKQIDAQRADLGAVQNRFDNTINNLKNIGENVSAARGRIEDTDFAA ETANLTKNQVLQQAGTAILAQANQLPQSVLSLLR Sequence length 394 AA Subcellular Location Secreted Function Flagellin is the subunit protein which polymerizes to form the filaments of bacterial flagella. It has been shown to have a role in the stimulation of inflammatory response via Toll-like receptor 5. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Ser/Thr) linked Experimentally Validated Glycosite(s) in Full Length Protein (Initiator Met removed) T190, S261 Experimentally Validated Glycosite(s ) in Mature Protein T189, S260 Glycosite(s) Annotated Protein Sequence >sp|P21184|FLICA_PSEAE A-type flagellin OS=Pseudomonas aeruginosa GN=fliC PE=1 SV=3 MALTVNTNIASLNTQRNLNNSSASLNTSLQRLSTGSRINSAKDDAAGLQIANRLTSQVNG LNVATKNANDGISLAQTAEGALQQSTNILQRMRDLSLQSANGSNSDSERTALNGEVKQLQ KELDRISNTTTFGGRKLLDGSFGVASFQVGSAANEIISVGIDEMSAESLNGTYFKADGGG AVTAATASGT*(190)VDIAIGITGGSAVNVKVDMKGNETAEQAAAKIAAAVNDANVGIGAFSDGD TISYVSKAGKDGSGAITSAVS*(261)GVVIADTGSTGVGTAAGVTPSATAFAKTNDTVAKIDIST AKGAQSAVLVIDEAIKQIDAQRADLGAVQNRFDNTINNLKNIGENVSAARGRIEDTDFAA ETANLTKNQVLQQAGTAILAQANQLPQSVLSLLR Sequence Around Glycosites (21 AA) GAVTAATASGTVDIAIGITGG DGSGAITSAVSGVVIADTGST Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Slower (and aberrant) migration on SDS-PAGE than its predicted mass, carbohydrate detection and deglycosylation with TFMS Technique(s) used for Glycosylated Residue(s) Detection LC-ES-MS/MS (liquid chromatography electrospray tandem mass spectrometry) analysis after β-elimination with ammonium hydroxide Protein Glycosylation- Implication Glycan Information Glycan Annotation Linkage: Rha-Ser/Thr. Heterogeneous glycan comprising up to 11 monosaccharide units that are O-linked through a rhamnose (deoxyhexose) residue; rhamnose, mannose, glucose, and 4-amino-4,6-dideoxyglucose (viosamine) as major components with novel, conserved trisaccharide capping. A single rhamnose residue is found at each site in P. aeruginosa JJ692. Technique(s) used for Glycan Identification Nanospray MS-MS analyses after HPLC of glycopeptide ions and GC-MS (gas chromatography-mass spectrometry) monosaccharide analysis Protein Glycosylation linked (PGL) gene(s) OST Gene Name pilO (tfpO) OST NCBI Gene ID OST GenBank Gene Sequence OST Protein Name PilO (TfpO) OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) 2.4.1.119 OST Genome Context Characterized Accessory Gene(s) Glycosyltransferases OrfA and OrfN are encoded in the glycosylation island (long in PAK strain and short in JJ692 strain). These are involved in the addition of the heterologous glycan and the proximal rhamnose residue, respectively. A cluster of 14 genes (OrfA to OrfN) constitute the glycosylation island (GI) that is essential for flagellin glycosylation. PGL Additional Links CAZy Literatures Reference(s) 1) Miller, W.L., Matewish, M.J., McNally, D.J., Ishiyama, N., Anderson, E.M., Brewer, D., Brisson, J.R., Berghuis, A.M. and Lam, J.S. (2008) Flagellin glycosylation in Pseudomonas aeruginosa PAK requires the O-antigen biosynthesis enzyme WbpO. J Biol Chem, 283, 3507-3518. [PubMed: 18065759] 2) Arora, S.K., Wolfgang, M.C., Lory, S. and Ramphal, R. (2004) Sequence polymorphism in the glycosylation island and flagellins of Pseudomonas aeruginosa. J Bacteriol, 186, 2115-2122. [PubMed: 15028697] Additional Comments Post translational modification was detected in the year 1990. It was confirmed as glycosylation in 1998. Year of Identification 1998 Year of Validation 2004