| ProGlyProt ID | BC167 |
| Organism Information |
| Organism Name | Pseudomonas aeruginosa Pa5196 |
| Domain | Bacteria |
| Classification | Family: Pseudomonadaceae
Order: Pseudomonadales
Class: Gammaproteobacteria
Division or phylum: "Proteobacteria" |
| Taxonomic ID (NCBI) | 287 |
| Genome Sequence (s) |
| EMBL | AY112720 |
| Gene Information |
| Gene Name | pilA |
| NCBI Gene ID | |
| Protein Information |
| Protein Name | PilA (Type IV pilin) |
| UniProtKB/SwissProt ID | Q8KQ32 |
| NCBI RefSeq | |
| EMBL-CDS | AAM52059.1 |
| 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 |
| Subcellular Location | Surface |
| 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. |
| Protein Structure |
| PDB ID | |
| Glycosylation Status |
| 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
AWPTLVAPTATPGAGQLNATL
QLNATLVGKYSSVDSTIASGY
LNATLVGKYSSVDSTIASGYP
TLVGKYSSVDSTIASGYPNGQ
KYSSVDSTIASGYPNGQITVT
|
| Glycosite Sequence Logo | seqlogo |
| Glycosite Sequence Logo |  |
| Technique(s) used for Glycosylation Detection | Slower migration on SDS-PAGE than its predicted mass and staining with GlycoProï¬le III fluorescent-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. |
| 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 |
| OST NCBI Gene ID | |
| OST GenBank Gene Sequence | |
| OST Protein Name | TfpW (arabinosyltransferase) |
| OST UniProtKB/ SwissProt ID | Q8KQ31 |
| OST NCBI RefSeq | |
| OST EMBL-CDS | AAM52060.1 |
| OST UniProtKB Sequence | >tr|Q8KQ31|Q8KQ31_PSEAE Putative arabinosyltransferase TfpW OS=Pseudomonas aeruginosa PE=4 SV=1
MKSLWVYLQGGGKKNVLLWVGVLCAVVWGLYGRALFFGYVWDDSILFLDKTDLLNSSISW
KVLSEPVLPGTSYLRPLVFLSFFVEFQIFGQSPFVSHAVNIAIFNLNVLLVFWLGSTLAR
HTERKYGLLLAFLAALFYAMHPALIESTVWASGRFDLLTTFFILAGLVVYLADIKSTVLR
AFLLCLCMAGALLSKELGIVFPVLLLCLWLAKECDCSTQRGGALLVQALFRNGWMLVALI
ITAFGYLVVRKHSAGGIYHSPVGLSYIADVVLGQRLPLETLKFYLLQSFLPFQSINVSHP
LNEVFERSVYVLLAGQMVVLILVLTVIYQALYRRSAAAWMFFAYLVCLLPVLHIIPLTIG
ENLGHERFLTAPLAFLALALVFVRYDRVLSKVGLSGAPLLRVGLVAGLLWLGMAGFTIYS
ILPMWTSDASLWAWAYKTHPESKYARYDYLYGALRGGYVELLDKEVSRLQKKYGGLEVPE
QILYAHRLISAGDSEGLNYLEGVLQVLPRFHDAENGRVALNDFKHLSSMQIGGAYAVYAE
GLSMLRGDAERALKYNKIAEWYLATGEVIPLMYQRSAILYALGDYGQAARIYQDQEGLHY
YMSQAVKKDMKRLVEGFCKEKGFPSDPCGELRERGVISKESEARQG
|
| OST EC Number (BRENDA) | |
| OST Genome Context | |
| Characterized Accessory Gene(s) | |
| PGL Additional Links | CAZy |
| Literatures |
| Reference(s) | 1) 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]
2) 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 |
| Additional Comments | It is the first report where ETD-MS has been used for bacetrial glycoprotein characterization.
It is also the ï¬rst report of glycosylation of a bacterial protein with a homooligosaccharide.
Group I P. aeruginosa pilins are glycosylated on a C-terminal Ser residue unlike Neisseria pilin which is glycosylated at a flexible-loop region within the globular domain of the protein.
TfpW (arabinosyltransferase) is the member of the GT-C glycosyltransferase family. Its role was determi |
| Year of Identification | 2004 |
| Year of Validation | 2007 |
