| 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 | 22701 |
| GlyTouCan | G26252RJ |
| 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) |
| Characterized Accessory Gene(s) | Four rhamnosyltransferases are encoded in the slg gene cluster. WsaC and WsaD are both α-1,3-rhamnosyltransferases, WsaE is α-1,2-rhamnosyltransferase, α-1,3-rhamnosyltransferase, 2-O-methyltransferase, and wsaF is α-1,2-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 |
| Year of Identification | 1984 |
| Year of Identification Month Wise | 1984 |
| Year of Validation | 2002 |
| Reference | Messner, P., Steiner, K., Zarschler, K. and Schäffer, C., 2008. S-layer nanoglycobiology of bacteria. Carbohydrate Research, 343(12), pp.1934-1951. |
| Corresponding Author | Christina Schaffer
Paul Messner |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria.
University of Natural Resources and Life Sciences Vienna, Center for NanoBiotechnology A-1180 Vienna, Gregor-Mendel-Strasse 33, Austria.
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| Reference | Steiner, K., Novotny, R., Werz, D.B., Zarschler, K., Seeberger, P.H., Hofinger, A., Kosma, P., Schäffer, C. and Messner, P., 2008. Molecular basis of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus. Journal of Biological Chemistry, 283(30), pp.21120-21133. |
| Corresponding Author | Christina Schaffer |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria. |
| Reference | Steiner, K., Novotny, R., Patel, K., Vinogradov, E., Whitfield, C., Valvano, M.A., Messner, P. and Schäffer, C., 2007. Functional characterization of the initiation enzyme of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus NRS 2004/3a. Journal of bacteriology, 189(7), pp.2590-2598. |
| Corresponding Author | Christina Schaffer |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria. |
| Reference | Steiner, K., Pohlentz, G., Dreisewerd, K., Berkenkamp, S., Messner, P., Peter-Katalinic, J. and Schäffer, C., 2006. New insights into the glycosylation of the surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a. Journal of bacteriology, 188(22), pp.7914-7921. |
| Corresponding Author | Christina Schaffer |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria. |
| Reference | Novotny, R., Schäffer, 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(4), pp.953-965. |
| Corresponding Author | Christina Schaffer |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria. |
| Reference | Schäffer, C., Wugeditsch, T., Kählig, 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. Journal of Biological Chemistry, 277(8), pp.6230-6239. |
| Corresponding Author | Christina Schaffer |
| Contact | Center for NanoBiotechnology, University of Natural Resources and Applied Life Sciences Vienna, A-1190 Vienna, Austria. |
