ProGP207

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ProGP ID ProGP207
Validation Status Characterized
Organism Information
Organism NameCampylobacter jejuni subsp. jejuni 81-176
Domain Bacteria
Classification Family: Campylobacteraceae
Order: Campylobacterales
Class: Epsilonproteobacteria
Division or phylum: "Proteobacteria"
Taxonomic ID (NCBI) 354242
Genome Sequence(s)
GenBank CP000538.1
EMBL CP000538
Organism Additional Information Campylobacter jejuni is a microaerophilic, Gram-negative, human pathogen that is the major cause of bacterial food-borne diarrhoea (gastroenteritis). It is most frequently responsible for a form of post-infection neuromuscular paralysis known as Guillain Barre' syndrome. It also leads to an immunoproliferative small intestine disease that is a rare malignant lymphoma of the intestine. Motility is essential for pathogenicity.
Gene Information
Gene NameflaA (CJJ81176_1339)
NCBI Gene ID 4682159
GenBank Gene Sequence 4682159
Protein Information
Protein NameFlagellin (FlaA)
UniProtKB/SwissProt ID Q2M5R2
NCBI RefSeq YP_001000997.1
EMBL-CDSEAQ72691.1
UniProtKB Sequence >tr|Q2M5R2|Q2M5R2_CAMJJ FlaA OS=Campylobacter jejuni subsp. jejuni serotype O:23/36 (strain 81-176) GN=flaA PE=4 SV=1 MGFRINTNVAALNAKANSDLNAKSLDASLSRLSSGLRINSAADDASGMAIADSLRSQANT LGQAISNGNDALGILQTADKAMDEQLKILDTIKTKATQAAQDGQSLKTRTMLQADINKLM EELDNIANTTSFNGKQLLSGNFTNQEFQIGASSNQTVKATIGATQSSKIGVTRFETGAQS FTSGVVGLTIKNYNGIEDFKFDNVVISTSVGTGLGALAEEINKSADKTGVRATYDVKTTG VYAIKEGTTSQEFAINGVTIGKIEYKDGDGNGSLISAINAVKDTTGVQASKDENGKLVLT SADGRGIKITGDIGVGSGILANQKENYGRLSLVKNDGRDINISGTNLSAIGMGTTDMISQ SSVSLRESKGQISATNADAMGFNSYKGGGKFVFTQNVSSISAFMSAQGSGFSRGSGFSVG SGKNLSVGLSQGIQIISSAASMSNTYVVSAGSGFSSGSGNSQFAALKTTAANTTDETAGV TTLKGAMAVMDIAETAITNLDQIRADIGSIQNQVTSTINNITVTQVNVKAAESQIRDVDF ASESANYSKANILAQSGSYAMAQANSSQQNVLRLLQ
Sequence length 576 AA
Subcellular LocationSecreted
Function It is the subunit protein which is polymerized into the flagellar filaments. Motility mediated by flagella is essential for virulence.
Glycosylation Status
Glycosylation Type O- (Ser) linked
Experimentally Validated Glycosite(s) in Full Length ProteinS207, S343, S348, T394, S398, S401, S405, S409, S418, S426, S430, S437, S441, S449, S452, S455, S458, S461, T482
Experimentally Validated Glycosite(s ) in Mature ProteinS206, S342, S347, T393, S397, S400, S404, S408, S417, S425, S429, S436, S440, S448, S451, S454, S457, S460, T481
Glycosite(s) Annotated Protein Sequence >tr|Q2M5R2|Q2M5R2_CAMJJ FlaA OS=Campylobacter jejuni subsp. jejuni serotype O:23/36 (strain 81-176) GN=flaA PE=4 SV=1 MGFRINTNVAALNAKANSDLNAKSLDASLSRLSSGLRINSAADDASGMAIADSLRSQANT LGQAISNGNDALGILQTADKAMDEQLKILDTIKTKATQAAQDGQSLKTRTMLQADINKLM EELDNIANTTSFNGKQLLSGNFTNQEFQIGASSNQTVKATIGATQSSKIGVTRFETGAQS FTSGVVGLTIKNYNGIEDFKFDNVVIS*(207)TSVGTGLGALAEEINKSADKTGVRATYDVKTTG VYAIKEGTTSQEFAINGVTIGKIEYKDGDGNGSLISAINAVKDTTGVQASKDENGKLVLT SADGRGIKITGDIGVGSGILANQKENYGRLSLVKNDGRDINIS*(343)GTNLS*(348)AIGMGTTDMISQ SSVSLRESKGQISATNADAMGFNSYKGGGKFVFT*(394)QNVS*(398)SIS*(401)AFMS*(405)AQGS*(409)GFSRGSGFS*(418)VGSGKNLS*(426)VGLS*(430) QGIQIIS*(437)SAAS*(441) MSNTYVVS*(449)AGS*(452) GFS*(455)SGS*(458) GNS*(461)QFAALKTTAANTTDETAGVTT*(482)LKGAMAVMDIAETAITNLDQIRADIGSIQNQVTSTINNITVTQVNVKAAESQIRDVDFASESANYSKANILAQSGSYAMAQANSSQQNVLRLLQ
Sequence Around Glycosites (21 AA) EDFKFDNVVISTSVGTGLGAL
VKNDGRDINISGTNLSAIGMG
RDINISGTNLSAIGMGTTDMI
SYKGGGKFVFTQNVSSISAFM
GGKFVFTQNVSSISAFMSAQG
FVFTQNVSSISAFMSAQGSGF
QNVSSISAFMSAQGSGFSRGS
SISAFMSAQGSGFSRGSGFSV
GSGFSRGSGFSVGSGKNLSVG
GFSVGSGKNLSVGLSQGIQII
GSGKNLSVGLSQGIQIISSAA
VGLSQGIQIISSAASMSNTYV
QGIQIISSAASMSNTYVVSAG
AASMSNTYVVSAGSGFSSGSG
MSNTYVVSAGSGFSSGSGNSQ
TYVVSAGSGFSSGSGNSQFAA
VSAGSGFSSGSGNSQFAALKT
GSGFSSGSGNSQFAALKTTAA
NTTDETAGVTTLKGAMAVMDI
Glycosite Sequence Logo
Technique(s) used for Glycosylation DetectionESMS (electrospray mass spectrometry); 10%- 6.5 kDa- mass excess detected
Technique(s) used for Glycosylated Residue(s) Detection A combination of μLC-ESMS (liquid chromatography-electrospray mass spectrometry) and MS-MS (tandem mass spectrometry) analyses; 10 out of 19 glycosylation sites were defined using nano-ESMS after base (NH4OH)-catalyzed β-elimination.
Protein Glycosylation- Implication Glycosylation is required for flagellar filament formation. Certain glycans mediate filament-filament interactions resulting in AAG (autoagglutination) and other glycans appear to be critical for structural subunit-subunit interactions within the filament. Modification with pseudaminic acid and derivatives is essential for targeting and/or secretion of flagellin. Also, specific structural modifications to the flagellin glycoform have been shown to be involved in the biological fitness of C. jejuni in colonization of chickens.
Glycan Information
Glycan Annotation Glycan represents 10% of the total mass of the protein.
Major glycan is pseudaminic acid and its derivatives, Pse5Pr7Pr, Pse5Ac7Ac8OAc, Pse5Am7Ac.
Pse5Ac7Ac (5,7-diacetamido-3,5,7,9 - tetradeoxy-L-glycero-L-manno- nonulosonic acid), with 5-acetamidino pseudaminic acid (Pse5Am7Ac) and 5,7-N-(2,3-dihydroxyproprionyl)-pseudaminic acid (Pse5Pr7Pr) are also present. In addition, novel glycans, Pse5Am7Ac8GlnAc and Pse5Ac7Ac8OAc, have also been found. S398 and S405 carry Pse5Pr7Pr moiety while T394, S401 and S409 are modified with Pse5Ac7Ac residues. Pse5Ac7Ac is also written as Pse5NAc7NAc (Pse). Microheterogeneity in glycosylation has been observed.
Technique(s) used for Glycan Identification Nano-ESMS and NMR analysis of HPLC fractions of trypsin digested glycopeptides including COSY(correlated spectroscopy) and NOESY (nuclear Overhauser effect spectroscopy).
Protein Glycosylation linked (PGL) gene(s)
OST ProGT IDProGT16
Characterized Accessory Gene(s)PseA, PseB, PseC, PseF, PseG, PseH, PseI are the enzymes required for CMP-Pse5NAc7NAc/CMP-Pse5NAc7Am production. PseB and PseC (dehydratase/aminotransferase) catalyze the first two steps of Pse5NAc7NAc (Pse) synthesis. PseI is the Pse synthase.
Additional CommentPseudaminic acid (Pse5Ac7Ac) has also been identified in the LPS of bacteria.
Pseudomonas pilin also carries pseudaminic acid like glycan (ProGlycProt ID BC166).
Sequon features: No specific sequence features except that all but one of total 19 O-glycosylated residues are restricted to the hydrophobic central, surface-exposed domain of flagellin when folded in the filament. It has been suggested that the local hydrophobicity upstream of Ser/Thr residues partially influences the site of glycosylation.
Glycosylation was detected in other strains by biotin-hydrazide labeling after periodate oxidation as early as 1996 (Ref. no. 7). It was the first report of eubacterial flagellin glycosylation.
Literature
Year of Identification2001
Year of Identification Month Wise2001.9.14
Year of Validation 2001
ReferenceMaita, N., Nyirenda, J., Igura, M., Kamishikiryo, J. and Kohda, D. (2010) Comparative structural biology of eubacterial and archaeal oligosaccharyltransferases. J Biol Chem, 285, 4941-4950. [PubMed: 20007322]
Author Maita, N., Nyirenda, J., Igura, M., Kamishikiryo, J. and Kohda, D.
Research GroupInstitute of Microbiology, Department of Biology, Swiss Federal Institute of Technology Zurich, ETH Hönggerberg, Switzerland.
Corresponding Author Kohda, D
ContactInstitute of Microbiology, Department of Biology, Swiss Federal Institute of Technology Zurich, ETH Hönggerberg, Switzerland.
ReferenceEwing, C.P., Andreishcheva, E. and Guerry, P. (2009) Functional characterization of flagellin glycosylation in Campylobacter jejuni 81-176. J Bacteriol, 191, 7086-7093. [PubMed: 19749047]
AuthorEwing CP1, Andreishcheva E, Guerry P.
Research GroupEnteric Diseases Department, Naval Medical Research Center, 503 Robert Grant Ave., Silver Spring, MD 20910, USA.
ContactEnteric Diseases Department, Naval Medical Research Center, 503 Robert Grant Ave., Silver Spring, MD 20910, USA.
ReferenceMcNally, D.J., Hui, J.P., Aubry, A.J., Mui, K.K., Guerry, P., Brisson, J.R., Logan, S.M. and Soo, E.C. (2006) Functional characterization of the flagellar glycosylation locus in Campylobacter jejuni 81-176 using a focused metabolomics approach. J Biol Chem, 281, 18489-18498. [PubMed: 16684771]
Author McNally, D.J., Hui, J.P., Aubry, A.J., Mui, K.K., Guerry, P., Brisson, J.R., Logan, S.M. Soo, E.C.
Research GroupNational Research Council, Institute for Biological Sciences, Ottawa, Ontario K1A 0R6, Canada.
Corresponding Author McNally DJ
ContactNational Research Council, Institute for Biological Sciences, Ottawa, Ontario K1A 0R6, Canada.
Reference 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]
Author Goon, S., Kelly, J.F., Logan, S.M., Ewing, C.P. Guerry, P.
Research GroupEnteric Diseases Department, Naval Medical Research Center, Silver Spring, MD 20910, USA.
Corresponding Author Guerry, P.
ContactEnteric Diseases Department, Naval Medical Research Center, Silver Spring, MD 20910, USA.
Reference 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]
Author Logan, S.M., Kelly, J.F., Thibault, P., Ewing, C.P. Guerry, P.
Research GroupInstitute for Biological Sciences, National Research Council, Ottawa, Canada.
Corresponding Author Guerry, P.
ContactInstitute for Biological Sciences, National Research Council, Ottawa, Canada.
Reference 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]
Author Thibault, P., Logan, S.M., Kelly, J.F., Brisson, J.R., Ewing, C.P., Trust, T.J. and Guerry, P
Research GroupInstitute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6, Canada.
Corresponding Author Guerry, P.
ContactInstitute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6, Canada.
Reference 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]
Author Doig, P., Kinsella, N., Guerry, P. and Trust, T.J.
Research GroupDepartment of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Corresponding Author Trust, T.J.
ContactDepartment of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.