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ProGlyProt ID
BC166
Organism Information
Organism Name
Pseudomonas aeruginosa 1244
Domain
Bacteria
Classification
Family: Pseudomonadaceae
Order: Pseudomonadales
Class: Gammaproteobacteria
Division or phylum: "Proteobacteria"
Taxonomic ID (NCBI)
 
 
Genome Sequence(s)
EMBL
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.
 
 
Gene Information
Gene Name
pilA
 
 
Protein Information
Protein Name
Fimbrial protein (pilin); (group I T4P)
UniProtKB/SwissProt ID
EMBL-CDS
UniProtKB Sequence
Sequence length
154 AA
Subcellular Location
Surface
Function
Structural unit of the pili which have a role in pathogenesis. They are required for colonization through adhesion and mediating surface translocation (twitching).
 
 
Protein Structure
Homology Model
Homology Model File
 
 
Glycosylation Status
Glycosylation Type
O (Ser) linked
Experimentally Validated Glycosite(s) in Full Length Protein
(Propeptide: 1-6) S154 (C-terminal residue)
Experimentally Validated Glycosite(s ) in Mature Protein
S148 (C-terminal residue)
Glycosite(s) Annotated Protein Sequence
>sp|P18774|FM12_PSEAE Fimbrial protein OS=Pseudomonas aeruginosa GN=pilA PE=1 SV
=1 MKAQKGFTLIELMIVVAIIGILAAIAIPQYQDYTARTQVTRAVSEVSALKTAAESAILEG KEIVSSATPK
DTQYDIGFTESTLLDGSGKSQIQVTDNQDGTVELVATLGKSSGSAIKGAV ITVSRKNDGVWNCKITKTPTAWKP
NYAPANCPKS*(154)
Sequence Around Glycosites (21 AA)
PNYAPANCPKS
Technique(s) used for Glycosylation Detection
DIG glycan detection (labeling with digoxigenin-succinyl-epsilon-amidocaproic acid hydrazide and anti-DIG antibody after periodate oxidation)
Technique(s) used for Glycosylated Residue(s) Detection
N-terminal sequencing and site-directed mutagenesis
Protein Glycosylation- Implication
Pilin glycosylation may function to protect the pilus against attack from proteolytic enzymes present as part of the host defense or as produced by P. aeruginosa itself. Presence of glycans, e.g., 5NβOHC47NfmPse on pilin fibrous structure would introduce a negative charge that would lower the pilus isoelectric point, influence solubility, and likely increase ionic interaction among pili and between the pili and extracellular structures thereby influencing pilus-dependent functions such as twitching motility and biofilm formation, processes that are important in pathogenesis as well.
 
 
Glycan Information
Glycan Annotation
Linkages: β-D-FucNAc-Ser.
A 666.5 Da trisaccharide containing pseudaminic acid, xylose, and N-acetylfucosamine [α-5NβOHC4 7NFmPse-(2→4)-β-Xyl-(1→3)-β-D-FucNAc-(1→3)-β-Ser].
Protein glycan molar ratio is 1:1.
It is structurally identical to the O-antigen repeating unit of P. aeruginosa serotype O7 LPS. This fact together with mutation studies suggest that the metabolic source of the pilin glycan is the O-antigen biosynthetic pathway. The pilin glycan differs from the O-antigen only in that the FucNAc residue is not O-acetylated.
BCSDB ID
Technique(s) used for Glycan Identification
The gradient NMR spectrum, tandem MS/MS analysis, and methylation analysis provided information on linkage and the sequence of oligosaccharide components. Xylosyl linkage was determined by GLC-MS analysis of partially methylated alditol acetate.
 
 
Protein Glycosylation linked (PGL) gene(s)
OST Gene Name
pilO (tfpO)
OST Protein Name
PilO (TfpO)
OST EC Number (BRENDA)
Characterized Accessory Gene(s)
WbpM, WbpL are encoded in the O-antigen biosynthesis cluster. WbpM is a UDP-GlcNAc C6 dehydratase/C4 reductase involved in the UDP-FucNAc biosynthesis. WbpL is a glycosyltransferase that transfers FucNAc from UDP-FucNAc to undecaprenol phosphate.
 
 
Literature
Reference(s)
1) Horzempa, J., Dean, C.R., Goldberg, J.B. and Castric, P. (2006) Pseudomonas aeruginosa 1244 pilin glycosylation: glycan substrate recognition. J Bacteriol, 188, 4244-4252. [PubMed: 16740931]
2) Smedley, J.G., 3rd, Jewell, E., Roguskie, J., Horzempa, J., Syboldt, A., Stolz, D.B. and Castric, P. (2005) Influence of pilin glycosylation on Pseudomonas aeruginosa 1244 pilus function. Infect Immun, 73, 7922-7931. [PubMed: 16299283]
3) Comer, J.E., Marshall, M.A., Blanch, V.J., Deal, C.D. and Castric, P. (2002) Identification of the Pseudomonas aeruginosa 1244 pilin glycosylation site. Infect Immun, 70, 2837-2845. [PubMed: 12010970]
4) DiGiandomenico, A., Matewish, M.J., Bisaillon, A., Stehle, J.R., Lam, J.S. and Castric, P. (2002) Glycosylation of Pseudomonas aeruginosa 1244 pilin: glycan substrate specificity. Mol Microbiol, 46, 519-530. [PubMed: 12406226]
5) Castric, P., Cassels, F.J. and Carlson, R.W. (2001) Structural characterization of the Pseudomonas aeruginosa 1244 pilin glycan. J Biol Chem, 276, 26479-26485. [PubMed: 11342554]
6) Castric, P. (1995) pilO, a gene required for glycosylation of Pseudomonas aeruginosa 1244 pilin. Microbiology, 141 ( Pt 5), 1247-1254. [PubMed: 7773418]
Additional Comments
O-antigen sugars are not sequentially added to the pilin. wbpM and wbpL are essential for the initial steps of O-antigen biosynthesis.
5-N-3 hydroxybutyryl-7-N-formylpseudaminic acid is a part of a trisaccharide modification on P. aeruginosa pilin. It is the second example of glycoprotein with pseudaminic acid containing glycan apart from Campylobacter flagellin protein.
The substrate recognition features required for catalysis by PilO enzyme have been shown to be present in the first (reducing) sugar, β-D-FucNAc.
Year of Identification
1995
Year of Validation
2002
 
 
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