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ProGlyProt ID
BC115
Organism Information
Organism Name
Campylobacter coli VC 167
Domain
Bacteria
Classification
Family: Campylobacteraceae
Order: Campylobacterales
Class: Epsilonproteobacteria
Division or phylum: "Proteobacteria"
Taxonomic ID (NCBI)
 
 
Genome Sequence(s)
EMBL
Organism Additional Information
Campylobacter sp. are known to cause bacterial food-borne gastroenteritis. Motility is essential for pathogenicity.
 
 
Gene Information
Gene Name
flaA
 
 
Protein Information
Protein Name
Flagellin A
UniProtKB/SwissProt ID
EMBL-CDS
UniProtKB Sequence
Sequence length
573 AA
Subcellular Location
Secreted
Function
It is the subunit protein which is polymerized into the flagellar filaments. Motility mediated by flagella is essential for virulence.
 
 
Protein Structure
Homology Model
Homology Model File
 
 
Glycosylation Status
Glycosylation Type
O (Ser) linked
Experimentally Validated Glycosite(s) in Full Length Protein
(Initiator Met removed) S207, S395, S396, S402, S409, S415, S423, S427, S434, S438, S440, S446, S452
Experimentally Validated Glycosite(s ) in Mature Protein
S206, S394, S395, S401, S408, S414, S422, S426, S433, S437, S439, S445, S451
Glycosite(s) Annotated Protein Sequence
>sp|P27053|FLAA_CAMCO Flagellin A OS=Campylobacter coli GN=flaA PE=3 SV=3 MGFRI
NTNVAALNAKANSDLNSRALDQSLSRLSSGLRINSAADDASGMAIADSLRSQANT LGQAISNGNDALGILQTAD
KAMDEQLKILDTIKTKATQAAQDGQSLKTRTMLQADINRLM EELDNIANTTSFNGKQLLSGGFTNQEFQIGSSS
NQTIKASIGATQSSKIGVTRFETGSQS FSSGTVGLTIKNYNGIEDFKFDSVVIS*(207)TSVGTGLGALAE
EINRNADKTGIRATFDVKSVG AYAIKAGNTSQDFAINGVVIGKVDYSDGDENGSLISAINAVKDTTGVQASKDE
NGKLVLT SADGRGIKITGSIGVGAGILHTENYGRLSLVKNDGRDINISGTGLSAIGMGATDMISQSS VSLRE
SKGQISAANADAMGFNAYNGGGAKQIIFAS*(395)S*(396)IAGFMS*(402)QAGSGFS*(409)AG
SGFS*(415)VGSGK NYS*(423)AILS*(427)ASIQIVS*(434)SARS*(438)IS*(44
0)
STYVVS*(446)TGSGFS*(452)AGSGNSQFAALRISTVSAHDETAGVTTL KGAMAVMDIAETAIT
NLDQIRADIGSVQNQITSTINNITVTQVNVKSAESQIRDVDFASE SANYSKANILAQSGSYAMAQANSSQQNVLRLLQ
Sequence Around Glycosites (21 AA)
EDFKFDSVVISTSVGTGLGAL
GGGAKQIIFASSIAGFMSQAG
GGAKQIIFASSIAGFMSQAGS
IFASSIAGFMSQAGSGFSAGS
GFMSQAGSGFSAGSGFSVGSG
GSGFSAGSGFSVGSGKNYSAI
GFSVGSGKNYSAILSASIQIV
GSGKNYSAILSASIQIVSSAR
AILSASIQIVSSARSISSTYV
ASIQIVSSARSISSTYVVSTG
IQIVSSARSISSTYVVSTGSG
ARSISSTYVVSTGSGFSAGSG
TYVVSTGSGFSAGSGNSQFAA
Glycosite Sequence Logo
Glycosite Sequence Logo
Technique(s) used for Glycosylation Detection
Biotin-hydrazide labeling after periodate oxidation, sialic acid-specific lectin LFA (limax flavus agglutinin)-binding, mass excess (approx. 6 kDa) detected by intact mass analysis using ESMS (electrospray mass spectrometry)
Technique(s) used for Glycosylated Residue(s) Detection
Edman sequencing
Protein Glycosylation- Implication
Sone level of glycosylation is necessary for filament assembly. Modification with either Pse5Ac7Ac or PseAm is sufficient for filament assembly.Modification with pseudaminic acid and derivatives is essential for targeting and/or secretion of flagellin. Glycosylation may be required for interaction of the subunits with the flagellin chaperone, FliS, with other components of the flagellar structure or in subunit interactions.
 
 
Glycan Information
Glycan Annotation
More Pse5Ac7Ac (5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid) in this case than from 81-176. Acetamidino and N-methylacetimidoyl derivatives of legionaminic acid, 315 Da Leg5Am7Ac (5-acetamidino-7-acetamido-3,5,7,9,-tetradeoxy-D-glycero-D-galacto-nonulosonic acid) and Leg5AmNMe7Ac (5-E/Z-N-(N-methylacetimidoyl)-7-acetamidino-3,5,7,9-tetradeoxy-D-glycero-D-galacto-nonulosonic), are also present. Leg5Ac7Ac (Leg) is legionaminic acid and Pse5Ac7Ac (Pse) is pseudaminic acid.
BCSDB ID
Technique(s) used for Glycan Identification
Homo- and heteronuclear correlated two-dimensional 1H NMR, 13C HSQC (heteronuclear single quantum coherence), HMBC (heteronuclear multiple bond coherence), COSY (correlated spectroscopy), TOCSY (total correlation spectroscopy), and NOESY (nuclear Overhauser effect spectroscopy).
 
 
Protein Glycosylation linked (PGL) gene(s)
Characterized Accessory Gene(s)
Products of ptmA-H genes are involved in the biosynthesis of Leg and Pse sugars that modify the flagellin.
Predicted Accessory Gene(s)
ptmC, ptmB, ptmG, and ptmH may encode a Leg5Ac7Ac synthase, a CMP-Leg5Ac7Ac synthetase, a CMP-Leg5Am7Ac acetamidino synthase, and an acetamidino N-methyltransferase, respectively. These genes have been shown to be involved in the Leg biosynthesis pathway.
 
 
Literature
Reference(s)
1) Logan, S.M., Hui, J.P., Vinogradov, E., Aubry, A.J., Melanson, J.E., Kelly, J.F., Nothaft, H. and Soo, E.C. (2009) Identification of novel carbohydrate modifications on Campylobacter jejuni 11168 flagellin using metabolomics-based approaches. FEBS J, 276, 1014-1023. [PMID: 19154343]
2) McNally, D.J., Aubry, A.J., Hui, J.P., Khieu, N.H., Whitfield, D., Ewing, C.P., Guerry, P., Brisson, J.R., Logan, S.M. and Soo, E.C. (2007) Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans. J Biol Chem, 282, 14463-14475. [PubMed: 17371878]
3) Goon, S., Kelly, J.F., Logan, S.M., Ewing, C.P. and Guerry, P. (2003) Pseudaminic acid, the major modification on Campylobacter flagellin, is synthesized via the Cj1293 gene. Mol Microbiol, 50, 659-671. [PubMed: 14617187]
4) Logan, S.M., Kelly, J.F., Thibault, P., Ewing, C.P. and Guerry, P. (2002) Structural heterogeneity of carbohydrate modifications affects serospecificity of Campylobacter flagellins. Mol Microbiol, 46, 587-597. [PubMed: 12406231]
5) Thibault, P., Logan, S.M., Kelly, J.F., Brisson, J.R., Ewing, C.P., Trust, T.J. and Guerry, P. (2001) Identification of the carbohydrate moieties and glycosylation motifs in Campylobacter jejuni flagellin. J Biol Chem, 276, 34862-34870. [PubMed: 11461915]
6) Doig, P., Kinsella, N., Guerry, P. and Trust, T.J. (1996) Characterization of a post-translational modification of Campylobacter flagellin: identification of a sero-specific glycosyl moiety. Mol Microbiol, 19, 379-387. [PubMed: 8825782]
7) Guerry, P., Doig, P., Alm, R.A., Burr, D.H., Kinsella, N. and Trust, T.J. (1996) Identification and characterization of genes required for post-translational modification of Campylobacter coli VC167 flagellin. Mol Microbiol, 19, 369-378. [PubMed: 8825781]
8) Logan, S.M., Trust, T.J. and Guerry, P. (1989) Evidence for posttranslational modification and gene duplication of Campylobacter flagellin. J Bacteriol, 171, 3031-3038. [PubMed: 2722741]
Additional Comments
It is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein.
Sequon feature: Consensus sequon does not appear to be related to the site of glycosylation. However, it has been suggested that the local hydrophobicity upstream of Ser/Thr residues partially influences the site of glycosylation.
It is the first report of eubacterial flagellin glycosylation. The modified residues were identified in 1989 using Edman sequencing (Ref. no. 8) but were not known to be glycosylated by then. Glycosyl residues on some of these sites were first identified in 2001 (Ref. no. 5).
Year of Identification
1996
Year of Validation
1989
 
 
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