|ProGP ID||ProGP230 (PilA (Type IV pilin))|
|Organism Name||Pseudomonas aeruginosa Pa5196|
|Classification|| Family: Pseudomonadaceae|
Division or phylum: "Proteobacteria"
|Taxonomic ID (NCBI)||287|
|Organism Additional Information||This Gram-negative opportunistic pathogen is responsible for nosocomial pneumonia. It possesses a multitude of virulence factors including type IV pili that mediate adhesion to host cells. It is also the major cause of mortality among cystic fibrosis (CF) patients.|
|Protein Name||PilA (Type IV pilin)|
|UniProtKB Sequence||>tr|Q8KQ32|Q8KQ32_PSEAE Type IV pilin structural subunit OS=Pseudomonas aeruginosa GN=pilA PE=3 SV=1 MKAQKGFTLIELMIVVAIIGILAAVAIPAYQDYITRGQVTEAVSLGGGLKSPLAEYGADK NAWPTLVAPTATPGAGQLNATLVGKYSSVDSTIASGYPNGQITVTMTQGKASGKKLTFST QDGGSSWACGNASIDGFAGTGTTIDAKYLPNACKP|
|Sequence length||155 AA|
|Function||Structural unit of the pili that have a role in adherence (colonization of both living and non-living surfaces) and twitching motility. These colonization factors (type IV pili) are also involved in biofilm formation.|
|Glycosylation Type||O- (Ser/Thr) linked|
|Experimentally Validated Glycosite(s) in Full Length Protein||(Propeptide: 1-6) T70, T72, S87, S88, S91, S95|
|Experimentally Validated Glycosite(s ) in Mature Protein||T64, T66, S81, S82, S85, S89|
|Glycosite(s) Annotated Protein Sequence||>tr|Q8KQ32|Q8KQ32_PSEAE Type IV pilin structural subunit OS=Pseudomonas aeruginosa GN=pilA PE=3 SV=1 MKAQKGFTLIELMIVVAIIGILAAVAIPAYQDYITRGQVTEAVSLGGGLKSPLAEYGADK NAWPTLVAPT*(70)AT*(72)PGAGQLNATLVGKYS*(87)S*(88)VDS*(91)TIAS*(95)GYPNGQITVTMTQGKASGKKLTFST QDGGSSWACGNASIDGFAGTGTTIDAKYLPNACKP|
|Sequence Around Glycosites (21 AA)|| KNAWPTLVAPTATPGAGQLNA
|ProGP Web Logo||Technique(s) used for Glycosylation Detection||Slower migration on SDS-PAGE than its predicted mass and staining with GlycoPro?le III ?uorescent-glycoprotein detection kit ||Technique(s) used for Glycosylated Residue(s) Detection || Validation of T70 and T72 glycosylated sites using site-directed mutagenesis and ETD-MS (electron transfer dissociation mass spectrometry). Rest of the sites were determined by a combination of nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) and nESI-MS/MS with front-end-collision induced dissociation (nESI-feCID-MS/MS).||Protein Glycosylation- Implication || Modifications of αβ-loop residues Thr64 and Thr66 are important for normal pilus assembly or extension/retraction dynamics. The Thr64Ala and Thr66Ala single and double mutants, as well as the nonglycosylated tfpW mutant, had fewer surface pili and reduced motility compared with the wild type.|| Glycan Information ||Glycan Annotation|| Linkages: FucNAc-Ser.|
Unusual homooligomers of α-1,5-linked D-arabinofuranose (α-1,5-D-Araf). Oligomer length varies from 3 to 8 units with an average length of 6 units. Trisaccharides of α-1,5-D-Araf are the principal modifications at Thr64 and Thr66, with additional mono- and disaccharides identified on Ser residues.
The pilin glycan is antigenically and chemically identical to that of Mycobacterium.
Microheterogeneity in glycosylation is observed.
|BCSDB ID||20019||Technique(s) used for Glycan Identification|| NMR spectroscopy including 1H-13C HSQC (heteronuclear single-quantum coherence) and 1H-13C HMBC (heteronuclear multiple bond coherence) and, chirality of glycans identified by GC-MS (gas chromatography-mass spectrometry).|| Protein Glycosylation linked (PGL) gene(s) ||OST Gene Name||TfpW (arabinosyltransferase)||OST ProGT ID||ProGT26 (TfpW) || Literature ||Reference||Kus, J.V., Kelly, J., Tessier, L., Harvey, H., Cvitkovitch, D.G. and Burrows, L.L. (2008) Modification of Pseudomonas aeruginosa Pa5196 type IV Pilins at multiple sites with D-Araf by a novel GT-C family Arabinosyltransferase, TfpW. J Bacteriol, 190, 7464-7478. [PubMed: 18805982]||Author||Kus, J.V., Kelly, J., Tessier, L., Harvey, H., Cvitkovitch, D.G. and Burrows, L.L||Research Group||Department of Biological Sciences, Duquesne University, 600 Forbes Ave., Pittsburgh, PA 15282, USA.|| Corresponding Author ||Burrows, L.L.||Contact||Department of Biological Sciences, Duquesne University, 600 Forbes Ave., Pittsburgh, PA 15282, USA.||Reference|| Voisin, S., Kus, J.V., Houliston, S., St-Michael, F., Watson, D., Cvitkovitch, D.G., Kelly, J., Brisson, J.R. and Burrows, L.L. (2007) Glycosylation of Pseudomonas aeruginosa strain Pa5196 type IV pilins with mycobacterium-like alpha-1,5-linked d-Araf oligosaccharides. J Bacteriol, 189, 151-159. [PubMed: 17085575]||Author|| Voisin, S., Kus, J.V., Houliston, S., St-Michael, F., Watson, D., Cvitkovitch, D.G., Kelly, J., Brisson, J.R. Burrows, L.L. ||Research Group||Institute for Biological Sciences, National Research Council, Ottawa, Canada.|| Corresponding Author || Burrows, L.L. ||Contact||Institute for Biological Sciences, National Research Council, Ottawa, Canada.||Reference|| Kus, J.V., Tullis, E., Cvitkovitch, D.G. and Burrows, L.L. (2004) Significant differences in type IV pilin allele distribution among Pseudomonas aeruginosa isolates from cystic fibrosis (CF) versus non-CF patients. Microbiology, 150, 1315-1326. [PubMed: 15133094]||Author|| Kus, J.V., Tullis, E., Cvitkovitch, D.G. Burrows, L.L. ||Research Group||1 Centre for Infection and Biomaterials Research, Hospital for Sick Children Research Institute and Department of Surgery, University of Toronto, 7142A Elm Wing, 555 University Avenue, Toronto, ON, Canada M5G 1X8
2 Adult Cystic Fibrosis Clinic, St Michaels Hospital, Toronto, ON, Canada
3 Department of Microbiology, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada|| Corresponding Author ||Burrows LL||Contact||1 Centre for Infection and Biomaterials Research, Hospital for Sick Children Research Institute and Department of Surgery, University of Toronto, 7142A Elm Wing, 555 University Avenue, Toronto, ON, Canada M5G 1X8 2. Department of Surgery, University of Toronto, 7142A Elm Wing, 555 University Avenue, Toronto, ON, Canada M5G 1X8.|