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ProGlyProt ID
BC145
Organism Information
Organism Name
Geobacillus (Bacillus) stearothermophilus NRS 2004/3a
Domain
Bacteria
Classification
Family: Bacillaceae
Order: Bacillales
Class: Bacilli (or Firmibacteria)
Division or phylum: "Firmicutes"
Taxonomic ID (NCBI)
 
 
Genome Sequence(s)
EMBL
 
 
Gene Information
Gene Name
sgsE
 
 
Protein Information
Protein Name
S-layer glycoprotein SgsE
UniProtKB/SwissProt ID
EMBL-CDS
UniProtKB Sequence
>tr|Q8VTF1|Q8VTF1_BACST Surface layer glycoprotein SgsE OS=Bacillus stearothermo
philus GN=sgsE PE=3 SV=1 MDKKKAVKLATASAVAASAFVAANPHTSQAATDVATVVSQAKAQMKEAYYT
YSHTVTETG QFPDIKDVYAAYNKAKQAYANAVAVVNKAGGAKKDAYLADLQATYETYVFKANPKSGEAR VATYI
DAYNYATKLDKMRQELKAAVDAKDLKKAEELYHKISYELKTRTVILDRVYGQSTR ELLRSTFKADAQALRDSLIY
DITVAMKAREAQDAVKAGNLDKAKAALDQVNQYVSKVTDA FKAELQKAAQDAKAAYEAALPPKVESVTAVNAKTL
EIKFNKAVDAATVIDNKGTSDTSDD VVKATAITLKAIDDQVPVSTVKASLSDDKKTLKLVVDGAQFFTKRYVVDI
KNVKTLDGKD VPAYTTTIDTTDSVRPSVLSFSYADNGLTLKVKFSEPLASVGTVKLYDGTTEISVSPKFT AGDD
EMTINLASSSVPVNKDLTLKIFGAVDYNGNVINPNPAELTVKKTTVDITKPVVQSI EAVNTKTVKVTFSEKLLSA
PTIKIGGQTASVSVDSTGLVYTATLASALSKGVYAVEVSDY KDLAGNSGDAYTKVVQLKADNTAPKFVSSQVVKI
NGVEHLVLTFDEEVTTGSNITVVQSS DKYIDENNVLKAVGADLKTTSDNFKLYLPTDGKSKSVALNISSLPKGTY
TVTLPNGLVSD LADNPYAERKQITFVRGSDSLTTKPALDKDYDSNGVKADNNNELVFAFTQNLDASALNLS NFN
INGLTVTKAVFDGDTKHIRVTLAPGANTWTGTHVITISNIKNTSGLVMDTVTVNEYM KENVAPTFTATLTSADVI
RVDFSEPVANATISRALSANNFIVKVDGNVVTVSNVYEDNNA TNLVQGSKGYKTVYLKLQSPVTDLSKPITLSAT
DIVDVNQTGAITDNNVVGNNVSATVVN VAK
Sequence length
903 AA
Subcellular Location
Surface
 
 
Glycosylation Status
Glycosylation Type
O (Ser/Thr) linked
Experimentally Validated Glycosite(s) in Full Length Protein
T590, T620, S794
Experimentally Validated Glycosite(s ) in Mature Protein
T590, T620, S794
Glycosite(s) Annotated Protein Sequence
>tr|Q8VTF1|Q8VTF1_BACST Surface layer glycoprotein SgsE OS=Bacillus stearothermo
philus GN=sgsE PE=3 SV=1 MDKKKAVKLATASAVAASAFVAANPHTSQAATDVATVVSQAKAQMKEAYY
TYSHTVTETG QFPDIKDVYAAYNKAKQAYANAVAVVNKAGGAKKDAYLADLQATYETYVFKANPKSGEAR VA
TYIDAYNYATKLDKMRQELKAAVDAKDLKKAEELYHKISYELKTRTVILDRVYGQSTR ELLRSTFKADAQALRD
SLIYDITVAMKAREAQDAVKAGNLDKAKAALDQVNQYVSKVTDA FKAELQKAAQDAKAAYEAALPPKVESVTAV
NAKTLEIKFNKAVDAATVIDNKGTSDTSDD VVKATAITLKAIDDQVPVSTVKASLSDDKKTLKLVVDGAQFFTK
RYVVDIKNVKTLDGKD VPAYTTTIDTTDSVRPSVLSFSYADNGLTLKVKFSEPLASVGTVKLYDGTTEISVSPK
FT AGDDEMTINLASSSVPVNKDLTLKIFGAVDYNGNVINPNPAELTVKKTTVDITKPVVQSI EAVNTKTVKV
TFSEKLLSAPTIKIGGQTASVSVDSTGLVYTATLASALSKGVYAVEVSDY KDLAGNSGDAYTKVVQLKADNTAP
KFVSSQVVKINGVEHLVLTFDEEVTT*(590)GSNITVVQSS DKYIDENNVLKAVGADLKTT*(620)SD
NFKLYLPTDGKSKSVALNISSLPKGTYTVTLPNGLVSD LADNPYAERKQITFVRGSDSLTTKPALDKDYDSNGV
KADNNNELVFAFTQNLDASALNLS NFNINGLTVTKAVFDGDTKHIRVTLAPGANTWTGTHVITISNIKNTSGLV
MDTVTVNEYM KENVAPTFTATLTS*(794)ADVIRVDFSEPVANATISRALSANNFIVKVDGNVVTVSNVYE
DNNA TNLVQGSKGYKTVYLKLQSPVTDLSKPITLSATDIVDVNQTGAITDNNVVGNNVSATVVN VAK
Sequence Around Glycosites (21 AA)
LVLTFDEEVTTGSNITVVQSS
LKAVGADLKTTSDNFKLYLPT
VAPTFTATLTSADVIRVDFSE
Glycosite Sequence Logo
Glycosite Sequence Logo
Technique(s) used for Glycosylation Detection
Periodic acid-Schiff (PAS) staining and carbohydrate content determination using orcinol-sulfuric acid assay
Technique(s) used for Glycosylated Residue(s) Detection
Edman degradation (of glycopeptides and deglycosylated peptides) and nano-ESI-QTOF-MS/MS (electrospray ionization-quadrupole time of flight tandem mass spectrometry)
 
 
Glycan Information
Glycan Annotation
Linkage: β-D-Galp-Ser/Thr.
[→2)-α-L-Rhap-(1→3)-β-L-Rhap-(1→2)-α-L-Rhap-(1→]n =13–18 (15 on average), with a 2-Omethyl group capping the terminal trisaccharide repeating unit at the non-reducing end of the glycan chains. The glycan chains are bound via the disaccharide (or trisaccharide with additional α-L-Rhap) core→3)-α-L-Rhap-(1→3)-α-L-Rhap-(1→and the linkage glycose β-D-Galp. Approx. 3.6% sugar content detected. Three inherently heterogeneic glycoprotein species of SgsE, 101.66 kDa, 108.68 kDa, and 115.73 kDa, have been determined. Nanoheterogeneity has been assigned to the S-layer glycans, with the most prevalent variation between 12 and 18 trisaccharide repeating units. One, two and three glycosylation sites, respectively, are occupied in these three glycoprotein species.
BCSDB ID
Technique(s) used for Glycan Identification
HPAEC/PAD (high performance anion-exchange chromatography/pulsed amperometric detection), MALDI-TOF MS analysis, 1D and 2D 1H and 13C NMR spectroscopy including 2D NOESY (nuclear Overhauser enhancement spectroscopy) and HMBC (heteronuclear multiple bond correlation spectroscopy)
 
 
Protein Glycosylation linked (PGL) gene(s)
OST Gene Name
wsaB
OST Protein Name
WsaB (putative oligosaccharyl:protein transferase)
Characterized Accessory Gene(s)
Four rhamnosyltransferases are encoded in the slg gene cluster. WsaC and WsaD are both α1,3-rhamnosyltransferases, WsaE is α1,2-rhamnosyltransferase, α1,3-rhamnosyltransferase, 2-O-methyltransferase, and wsaF is α1,2-rhamnosyltransferase. Membrane-anchored WsaD initiates the reaction by transfering activated β-L-Rha to the lipid-bound galactose primer. WsaP is UDP Gal:phosphoryl-polyprenol Gal-1-phosphate transferase.
 
 
Literature
Reference(s)
1) Messner, P., Steiner, K., Zarschler, K. and Schaffer, C. (2008) S-layer nanoglycobiology of bacteria. Carbohydr Res, 343, 1934-1951. [PubMed: 18336801]
2) Steiner, K., Novotny, R., Werz, D.B., Zarschler, K., Seeberger, P.H., Hofinger, A., Kosma, P., Schaffer, C. and Messner, P. (2008) Molecular basis of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus. J Biol Chem, 283, 21120-21133. [PubMed: 18515358]
3) Steiner, K., Novotny, R., Patel, K., Vinogradov, E., Whitfield, C., Valvano, M.A., Messner, P. and Schaffer, C. (2007) Functional characterization of the initiation enzyme of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus NRS 2004/3a. J Bacteriol, 189, 2590-2598. [PubMed: 17237178]
4) Steiner, K., Pohlentz, G., Dreisewerd, K., Berkenkamp, S., Messner, P., Peter-Katalinic, J. and Schaffer, C. (2006) New insights into the glycosylation of the surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a. J Bacteriol, 188, 7914-7921. [PubMed: 16963578]
5) Novotny, R., Schaffer, C., Strauss, J. and Messner, P. (2004) S-layer glycan-specific loci on the chromosome of Geobacillus stearothermophilus NRS 2004/3a and dTDP-L-rhamnose biosynthesis potential of G. stearothermophilus strains. Microbiology, 150, 953-965. [PubMed: 15073305]
6) Schaffer, C., Wugeditsch, T., Kahlig, H., Scheberl, A., Zayni, S. and Messner, P. (2002) The surface layer (S-layer) glycoprotein of Geobacillus stearothermophilus NRS 2004/3a. Analysis of its glycosylation. J Biol Chem, 277, 6230-6239. [PubMed: 11741945]
7) Christian, R., Schulz, G., Unger, F.M., Messner, P., Kupcu, Z. and Sleytr, U.B. (1986) Structure of a rhamnan from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a. Carbohydr Res, 150, 265-272. [PubMed: 3756958]
8) Kupcu, Z., Marz, L., Messner, P. and Sleytr, U.B. (1984) Evidence for the glycoprotein nature of the crystalline cell wall surface layer of Bacillus stearothermophilus strain NRS2004/3a. FEBS Lett, 173, 185-190. [PubMed: 6745426]
Year of Identification
1984
Year of Validation
2002
 
 
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