ProGlyProt ID BC145 Organism Information Organism Name Geobacillus stearoth Domain Bacteria Classification Family: Bacillaceae Order: Bacillales Class: Bacilli (or Firmibacteria) Division or phylum: "Firmicutes" Taxonomic ID (NCBI) 1422 Genome Sequence (s) EMBL AF328862 Gene Information Gene Name sgsE NCBI Gene ID Protein Information Protein Name S-layer glycoprotein SgsE UniProtKB/SwissProt ID Q8VTF1 NCBI RefSeq EMBL-CDS AAL46630.1 UniProtKB Sequence >tr|Q8VTF1|Q8VTF1_BACST Surface layer glycoprotein SgsE OS=Bacillus stearothermophilus GN=sgsE PE=3 SV=1 MDKKKAVKLATASAVAASAFVAANPHTSQAATDVATVVSQAKAQMKEAYYTYSHTVTETG QFPDIKDVYAAYNKAKQAYANAVAVVNKAGGAKKDAYLADLQATYETYVFKANPKSGEAR VATYIDAYNYATKLDKMRQELKAAVDAKDLKKAEELYHKISYELKTRTVILDRVYGQSTR ELLRSTFKADAQALRDSLIYDITVAMKAREAQDAVKAGNLDKAKAALDQVNQYVSKVTDA FKAELQKAAQDAKAAYEAALPPKVESVTAVNAKTLEIKFNKAVDAATVIDNKGTSDTSDD VVKATAITLKAIDDQVPVSTVKASLSDDKKTLKLVVDGAQFFTKRYVVDIKNVKTLDGKD VPAYTTTIDTTDSVRPSVLSFSYADNGLTLKVKFSEPLASVGTVKLYDGTTEISVSPKFT AGDDEMTINLASSSVPVNKDLTLKIFGAVDYNGNVINPNPAELTVKKTTVDITKPVVQSI EAVNTKTVKVTFSEKLLSAPTIKIGGQTASVSVDSTGLVYTATLASALSKGVYAVEVSDY KDLAGNSGDAYTKVVQLKADNTAPKFVSSQVVKINGVEHLVLTFDEEVTTGSNITVVQSS DKYIDENNVLKAVGADLKTTSDNFKLYLPTDGKSKSVALNISSLPKGTYTVTLPNGLVSD LADNPYAERKQITFVRGSDSLTTKPALDKDYDSNGVKADNNNELVFAFTQNLDASALNLS NFNINGLTVTKAVFDGDTKHIRVTLAPGANTWTGTHVITISNIKNTSGLVMDTVTVNEYM KENVAPTFTATLTSADVIRVDFSEPVANATISRALSANNFIVKVDGNVVTVSNVYEDNNA TNLVQGSKGYKTVYLKLQSPVTDLSKPITLSATDIVDVNQTGAITDNNVVGNNVSATVVN VAK Sequence length 903 AA Subcellular Location Surface Function Protein Structure PDB ID 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 stearothermophilus GN=sgsE PE=3 SV=1 MDKKKAVKLATASAVAASAFVAANPHTSQAATDVATVVSQAKAQMKEAYYTYSHTVTETG QFPDIKDVYAAYNKAKQAYANAVAVVNKAGGAKKDAYLADLQATYETYVFKANPKSGEAR VATYIDAYNYATKLDKMRQELKAAVDAKDLKKAEELYHKISYELKTRTVILDRVYGQSTR ELLRSTFKADAQALRDSLIYDITVAMKAREAQDAVKAGNLDKAKAALDQVNQYVSKVTDA FKAELQKAAQDAKAAYEAALPPKVESVTAVNAKTLEIKFNKAVDAATVIDNKGTSDTSDD VVKATAITLKAIDDQVPVSTVKASLSDDKKTLKLVVDGAQFFTKRYVVDIKNVKTLDGKD VPAYTTTIDTTDSVRPSVLSFSYADNGLTLKVKFSEPLASVGTVKLYDGTTEISVSPKFT AGDDEMTINLASSSVPVNKDLTLKIFGAVDYNGNVINPNPAELTVKKTTVDITKPVVQSI EAVNTKTVKVTFSEKLLSAPTIKIGGQTASVSVDSTGLVYTATLASALSKGVYAVEVSDY KDLAGNSGDAYTKVVQLKADNTAPKFVSSQVVKINGVEHLVLTFDEEVTT*(590)GSNITVVQSS DKYIDENNVLKAVGADLKTT*(620)SDNFKLYLPTDGKSKSVALNISSLPKGTYTVTLPNGLVSD LADNPYAERKQITFVRGSDSLTTKPALDKDYDSNGVKADNNNELVFAFTQNLDASALNLS NFNINGLTVTKAVFDGDTKHIRVTLAPGANTWTGTHVITISNIKNTSGLVMDTVTVNEYM KENVAPTFTATLTS*(794)ADVIRVDFSEPVANATISRALSANNFIVKVDGNVVTVSNVYEDNNA TNLVQGSKGYKTVYLKLQSPVTDLSKPITLSATDIVDVNQTGAITDNNVVGNNVSATVVN VAK Sequence Around Glycosites (21 AA) LVLTFDEEVTTGSNITVVQSS LKAVGADLKTTSDNFKLYLPT VAPTFTATLTSADVIRVDFSE Glycosite Sequence Logo seqlogo 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) Protein Glycosylation- Implication 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 o 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 NCBI Gene ID OST GenBank Gene Sequence OST Protein Name WsaB (putative oligosaccharyl:protein transferase) OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) OST Genome Context 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. PGL Additional Links CAZy Literatures 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., Additional Comments Year of Identification 1984 Year of Validation 2002