ProGP152 (Type A Flagellin)
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| ProGP ID | ProGP152 (Type A Flagellin) |
| Validation Status | Characterized |
| Organism Information | |
| Organism Name | Pseudomonas aeruginosa PAK/5939 |
| Domain | Bacteria |
| Classification | Phylum : Proteobacteria Class : Gammaproteobacteria Orders : Pseudomonadales Family : Pseudomonadaceae Genus : Pseudomonas Species : aeruginosa Strain : PAK/5939 |
| Taxonomic ID (NCBI) | 287 |
| Genome Information | |
| GenBank | M57501 |
| EMBL | M57501 |
| Organism Additional Information | It is a Gram-negative opportunistic pathogen that is responsible for nosocomial pneumonia. Its virulence factors include exotoxins, proteases, lipases, lipopolysaccharide (LPS), pili, and flagellin protein. |
| Gene Information | |
| Gene Name | fliC |
| Protein Information | |
| Protein Name | Type A Flagellin |
| UniProtKB/SwissProt ID | P21184 |
| 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. |
| 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 |
| 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 |
| 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. |
| BCSDB ID | 22702 |
| 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 ProGT ID | ProGT1 |
| 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. |
| Accessory Gene(s)Progt ID | ProGT1.1-ProGT1.2 |
| Additional Comment | Post translational modification was detected in the year 1990. It was confirmed as glycosylation in 1998. |
| Literature | |
| Year of Identification | 1998 |
| Year of Identification Month Wise | 1998.06.01 |
| Year of Validation | 2004 |
| Reference | 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. Journal of Biological Chemistry, 283(6), pp.3507-3518. |
| Corresponding Author | Joseph S Lam |
| Contact | Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada. |
| Reference | Arora, S.K., Wolfgang, M.C., Lory, S. and Ramphal, R., 2004. Sequence polymorphism in the glycosylation island and flagellins of Pseudomonas aeruginosa. Journal of bacteriology, 186(7), pp.2115-2122. |
| Corresponding Author | Reuben Ramphal |
| Contact | Department of Medicine/Infectious Diseases, University of Florida, Gainesville, FL 32610, USA |
| Reference | Schirm, M., Arora, S.K., Verma, A., Vinogradov, E., Thibault, P., Ramphal, R. and Logan, S.M., 2004. Structural and genetic characterization of glycosylation of type a flagellin in Pseudomonas aeruginosa. Journal of bacteriology, 186(9), pp.2523-2531. |
| Corresponding Author | Susan M Logan |
| Contact | Institute for Biological Sciences, National Research Council, Ottawa, Ontario, Canada |
| Reference | Arora, S.K., Bangera, M., Lory, S. and Ramphal, R., 2001. A genomic island in Pseudomonas aeruginosa carries the determinants of flagellin glycosylation. Proceedings of the National Academy of Sciences, 98(16), pp.9342-9347. |
| Corresponding Author | Reuben Ramphal |
| Contact | Department of Medicine/Infectious Diseases, University of Florida, Gainesville, FL 32610, USA |
| Reference | Brimer, C.D. and Montie, T.C., 1998. Cloning and comparison of fliC genes and identification of glycosylation in the flagellin of Pseudomonas aeruginosa a-type strains. Journal of bacteriology, 180(12), pp.3209-3217. |
| Corresponding Author | T. C. Montie |
| Contact | Department of Microbiology, University of Tennessee, Knoxville 37996-0845, USA. |
| Reference | Totten, P.A. and Lory, S.T.E.P.H.E.N., 1990. Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK. Journal of bacteriology, 172(12), pp.7188-7199. |
| Corresponding Author | S Lory |
| Contact | Department of Microbiology, School of Medicine, University of Washington, Seattle 98195. |