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ProGlyProt ID
AC114
Organism Information
Organism Name
Methanococcus voltae PS
Domain
Archaea
Classification
Family: Methanococcaceae
Order: Methanococcales
Class: Methanococci or Methanothermea
Division or phylum: "Euryarchaeota"
Taxonomic ID (NCBI)
 
 
Genome Sequence(s)
EMBL
 
 
Gene Information
Gene Name
flaB3
 
 
Protein Information
Protein Name
Fla B3
UniProtKB/SwissProt ID
EMBL-CDS
UniProtKB Sequence
Sequence length
239 AA
Subcellular Location
Surface associated
Function
Flagellin structural protein
 
 
Glycosylation Status
Glycosylation Type
N (Asn) linked
Experimentally Validated Glycosite(s) in Full Length Protein
(Propeptide: 1-11) N115, N128
Experimentally Validated Glycosite(s ) in Mature Protein
N104, N117
Glycosite(s) Annotated Protein Sequence
>sp|P27805|FLAB3_METVO Flagellin B3 OS=Methanococcus voltae GN=flaB3 PE=1 SV=1
MLKNFMKNKKGAVGIGTLIIFIALVLVAAVAASVIINTAGKLQHKAAVVGQETTQQVASG LQVVKITGHSVDQY
NLDKIAILVSPNIGDEIDLATTVVTFSTDDRKMSLLYDSSN*(115)ASNGG KVRLSTAN*(128)GTSD
IFKYDDVYAIGAWPFEDPTYGQSDQDPNEKKFGIVVLQDMDNSVSGEH PTVNYGDKVLLAINIGNIVGENIGNR
IRIQGEVVPEFGAPGIIDFTTPPVYANRVVALQ
Sequence Around Glycosites (21 AA)
RKMSLLYDSSNASNGGKVRLS
NGGKVRLSTANGTSDIFKYDD
Glycosite Sequence Logo
Glycosite Sequence Logo
Technique(s) used for Glycosylation Detection
Mass shift detected on SDS-polyacrylamide gel and glycan identification by nano-LC-MS/MS
Technique(s) used for Glycosylated Residue(s) Detection
Nano-LC-MS/MS (nano-liquid chromatography-electrospray tandem mass spectrometry)
Protein Glycosylation- Implication
The glycans on flagellin proteins are involved in the flagella assembly process.
 
 
Glycan Information
Glycan Annotation
Linkage: β-GlcNAc-Asn.
Trisaccharide (779 Da) composed of β-Manp NAcA6Thr-(1– 4)- β-Glcp NAc3NAcA-(1–3)-β-Glcp NAc. The sugars are mannuronic acid with the attached threonine, diacetylated glucuronic acid, N-acetylglucosamine.
In a second version of this strain (M. voltae PS*), the glycan is modified with one additional residue (either 220 or 262 Da) linked to the terminal modified mannuronic acid.
BCSDB ID
Technique(s) used for Glycan Identification
NanoLC–MS/MS analysis and NMR spectroscopy- COSY (correlated spectroscopy), TOCSY (total correlation spectroscopy), NOESY (nuclear Overhauser effect spectroscopy) spectra, and 1H-13C HMBC (heteronuclear multiple bond coherence) spectra.
 
 
Protein Glycosylation linked (PGL) gene(s)
OST Gene Name
aglB
OST Protein Name
AglB
OST UniProtKB/ SwissProt ID
OST EMBL-CDS
OST UniProtKB Sequence
>tr|Q2EMT4|Q2EMT4_METVO STT3 OS=Methanococcus voltae GN=MVO1749 PE=4 SV=1 MTENNE
KVKNSDSANNQSSKNSKFNFNFEDKKVKCAKTILIIIFLAFLSFQMRAQTADMG FTTNEQYLDVFSDDNGRMYLT
ALDPYYYLRMSENYLENGHTGDTLKNIDGQQVPWDSYKY GPTGARATFNLLSVVTVWVYQVWHAMDSTVTLMNAA
FWVPAILSMFLITPIFFTVRRITS SDIGGAVAAILASLSPSIFVKTVAGFSDTPILEILPLLFIVWFIIEAIHYS
KEKNYKSLI YGLLATLMLALYPFMWSAWWYGYYIVIAFLVIYAIYKGISYNSIAKYTKSKNNNHKDKIE SEKLE
MLNILKISGLFIIGGAVLITALYGVSTTMNALQAPLNYLGLDEVSSQTGWPNVLT TVSELDTASLDEIISSSLGS
IHLFAIGLIGIFLSLFRKVLTPVKQISNGLAEKLDIKYAL LLIIWFAVTFLAASKGVRFVALMVPPLSIGVGIFV
GFIEQFIKNNLDKKYEYVAYPTIAI IVLYALFTIYRADSADLVRMLLPSNYVPIAEGIMLASLAVLIIYKVAELI
AESNKKLVMN KIFMILLAIGLITPTIATIVPFYSVPTYNDGWGESLEWINTQTPNNSVVTCWWDNGHIYT WKTD
RMVTFDGSSQNTPRAYWVGRAFSTSNESLANGIFRMLASSGDKAYTTDSVLIKKTG SIKNTVDVLNEILPLTKSD
AQKALKNSSYKFTDTEVSEILDATHPKVTNPDYLITYNRMT SIASVWSYFGNWDFNLPAGTSRSEREAGSFQGLQ
TYATNINDTLIVRSLIQQTAEYNIYT LIEVRNETLTGAMMAVTNDGQMQTQQLNMHKVKLMVNENGKSKMYNSLA
DPDGQLSLLIK VDKNSIIGTDGSNNPVYSSSSWMATANLEDSVYSKLHFFDGEGLDTIKLEKESLDPTANG VQP
GFKVFSVDYGNYSK
Characterized Accessory Gene(s)
AglH, AglC, AglK, AglA glycosyltransferases are involved in the biosynthesis of the glycan. AglH is a GlcNAc-1-phosphate transferase transfering first sugar to dolichol pyrophosphate. AglC and AglK add the second sugar residue and AglA adds the third sugar to the glycan.
Characterized by gene deletion studies
PGL Additional Links
 
 
Literature
Reference(s)
1) Chaban, B., Logan, S.M., Kelly, J.F. and Jarrell, K.F. (2009) AglC and AglK are involved in biosynthesis and attachment of diacetylated glucuronic acid to the N-glycan in Methanococcus voltae. J Bacteriol, 191, 187-195. [PubMed: 18978056]
2) Shams-Eldin, H., Chaban, B., Niehus, S., Schwarz, R.T. and Jarrell, K.F. (2008) Identification of the archaeal alg7 gene homolog (encoding N-acetylglucosamine-1-phosphate transferase) of the N-linked glycosylation system by cross-domain complementation in Saccharomyces cerevisiae. J Bacteriol, 190, 2217-2220. [PubMed: 18178736]
3) Chaban, B., Voisin, S., Kelly, J., Logan, S.M. and Jarrell, K.F. (2006) Identification of genes involved in the biosynthesis and attachment of Methanococcus voltae N-linked glycans: insight into N-linked glycosylation pathways in Archaea. Mol Microbiol, 61, 259-268. [PubMed: 16824110]
4) Voisin, S., Houliston, R.S., Kelly, J., Brisson, J.R., Watson, D., Bardy, S.L., Jarrell, K.F. and Logan, S.M. (2005) Identification and characterization of the unique N-linked glycan common to the flagellins and S-layer glycoprotein of Methanococcus voltae. J Biol Chem, 280, 16586-16593. [PubMed: 15723834]
5) Bayley, D.P. and Jarrell, K.F. (1999) Overexpression of Methanococcus voltae flagellin subunits in Escherichia coli and Pseudomonas aeruginosa: a source of archaeal preflagellin. J Bacteriol, 181, 4146-4153. [PubMed: 10400569]
Additional Comments
Post translational modification was detected in the year 1999. It was confirmed as glycosylation (by identifying the glycans) in 2005.
Year of Identification
2005
Year of Validation
2005
 
 
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