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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)
 
 
Genome Sequence(s)
EMBL
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
EMBL-CDS
UniProtKB Sequence
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
Homology Model
Homology Model File
 
 
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 S
V=3 MALTVNTNIASLNTQRNLNNSSASLNTSLQRLSTGSRINSAKDDAAGLQIANRLTSQVNG LNVATKNAN
DGISLAQTAEGALQQSTNILQRMRDLSLQSANGSNSDSERTALNGEVKQLQ KELDRISNTTTFGGRKLLDGSFG
VASFQVGSAANEIISVGIDEMSAESLNGTYFKADGGG AVTAATASGT*(190)VDIAIGITGGSAVNVKVDM
KGNETAEQAAAKIAAAVNDANVGIGAFSDGD TISYVSKAGKDGSGAITSAVS*(261)GVVIADTGSTGVGT
AAGVTPSATAFAKTNDTVAKIDIST AKGAQSAVLVIDEAIKQIDAQRADLGAVQNRFDNTINNLKNIGENVSAA
RGRIEDTDFAA 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
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 Protein Name
PilO (TfpO)
OST EC Number (BRENDA)
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.
 
 
Literature
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]
3) 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. J Bacteriol, 186, 2523-2531. [PubMed: 15090491]
4) Arora, S.K., Bangera, M., Lory, S. and Ramphal, R. (2001) A genomic island in Pseudomonas aeruginosa carries the determinants of flagellin glycosylation. Proc Natl Acad Sci U S A, 98, 9342-9347. [PubMed: 11481492]
5) 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. J Bacteriol, 180, 3209-3217. [PubMed: 9620973]
6) Totten, P.A. and Lory, S. (1990) Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK. J Bacteriol, 172, 7188-7199. [PubMed: 2123866]
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
 
 
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