| Technique(s) used for Glycosylation Detection | Deglycosylation with anhydrous hydrogen fluoride. |
| Technique(s) used for Glycosylated Residue(s) Detection | Nano-LC-ES-MS/MS (nano-liquid chromatography-electrospray tandem mass spectrometry) |
| Protein Glycosylation- Implication | N-glycosylation endows H. volcanii with an ability to maintain an intact and stable cell envelope (correct S-layer architecture and stable association of the S-layer glycoprotein within the S-layer) in hypersaline surroundings, ensuring survival in this extreme environment. |
| Glycan Information |
| Glycan Annotation | Sulfoquinovose-hexose-based glycan, N-linked via a chitobiose core.
A trisaccharide corresponding to glucuronic acid (GlcA)-β-1,4-GlcA-β-1,4-glucose-β-1-Asn, a Tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-β→1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α→1,4-GlcA-β1,4-glucose-β1-Asn.
The hexose at the non-reducing end of the pentasaccharide is a mannose.
In high salt (3.4 M NaCl) conditions, N13 and N83 are modified by a pentasaccharide while dolicholphosphate is modified by a Tetrasaccharide comprising the first four pentasaccharide residues.
Cells grown at low salinity (1.75 M NaCl) possess dolichol phosphate and N498 residue (in S-layer glycoprotein) modified by a distinct Tetrasaccharide comprising a sulfated hexose, two hexoses and a rhamnose.
A novel glycan, N-acetylglucosamine2-hexose2-(sulfoquinovose-hexose)6, is bound to Asn-732. |
| GlyTouCan | G31587VS |
| Technique(s) used for Glycan Identification | LC-ESI-MS, MALDI-TOF MS, Nano-LC-MS/MS and NMR |
| Protein Glycosylation linked (PGL) gene(s) |
| OST Gene Name | AglB |
| OST ProGT ID | ProGT15 |
| Characterized Accessory Gene(s) | AglD, AglE, AglI, AglG and AglJ are glycosyltransferases involved in the pentasaccharide assembly. AglJ adds the first sugar of the glycan and AglD adds the last one. All except AglD are encoded in the agl gene island or cluster. The aglB is also present in this island.
Characterized by a combination of gene deletion, mass spectroscopy and biochemical characterization. |
| Accessory Gene(s)Progt ID | ProGT15.1, ProGT15.2, Pro |
| Additional Comment | Based on analogy to H. salinarium and presence of similar glycans, the S layer protein in H. volcanni that exhibits a cluster of threonine residues (yet uncharacterized) at C terminus of the protein nearby hydrophobic membrane anchor has been speculated to be glycosylated with glucosyl-(1-2)-galactose disaccharides. The probable glycosylated cluster sequence has been attributed to act as a spacer between membrane anchor and extracellular domain akin to periplasmic space in gram negative bacteria. |
| Literature |
| Year of Identification | 1990 |
| Year of Identification Month Wise | 1990.12.25 |
| Year of Validation | 1992 |
| Reference | Kandiba, L., Lin, C.W., Aebi, M., Eichler, J. and Guerardel, Y., 2016. Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. Glycobiology, 26(7), pp.745-756. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Parente, J., Casabuono, A., Ferrari, M.C., Paggi, R.A., De Castro, R.E., Couto, A.S. and Giménez, M.I., 2014. A rhomboid protease gene deletion affects a novel oligosaccharide N-linked to the S-layer glycoprotein of Haloferax volcanii. Journal of Biological Chemistry, 289(16), pp.11304-11317. |
| Corresponding Author | María Inés Giménez |
| Contact | The Institute of Biological Research, National University of Mar del Plata-National Council of Scientific and Technical Research, Funes 3250 4th level, 7600 Mar del Plata, Province of Buenos Aires, Argentina |
| Reference | Guan, Z., Naparstek, S., Calo, D. and Eichler, J., 2012. Protein glycosylation as an adaptive response in Archaea: growth at different salt concentrations leads to alterations in Haloferax volcanii S‐layer glycoprotein N‐glycosylation. Environmental microbiology, 14(3), pp.743-753. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Kaminski, L., Abu-Qarn, M., Guan, Z., Naparstek, S., Ventura, V.V., Raetz, C.R., Hitchen, P.G., Dell, A. and Eichler, J., 2010. AglJ adds the first sugar of the N-linked pentasaccharide decorating the Haloferax volcanii S-layer glycoprotein. Journal of bacteriology, 192(21), pp.5572-5579. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Kaminski, L. and Eichler, J., 2010. Identification of residues important for the activity of Haloferax volcanii AglD, a component of the archaeal N-glycosylation pathway. Archaea, 2010. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Magidovich, H., Yurist‐Doutsch, S., Konrad, Z., Ventura, V.V., Dell, A., Hitchen, P.G. and Eichler, J., 2010. AglP is a S‐adenosyl‐l‐methionine‐dependent methyltransferase that participates in the N‐glycosylation pathway of Haloferax volcanii. Molecular microbiology, 76(1), pp.190-199. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Yurist-Doutsch, S. and Eichler, J., 2009. Manual annotation, transcriptional analysis, and protein expression studies reveal novel genes in the agl cluster responsible for N glycosylation in the halophilic archaeon Haloferax volcanii. Journal of bacteriology, 191(9), pp.3068-3075. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Abu-Qarn, M., Giordano, A., Battaglia, F., Trauner, A., Hitchen, P.G., Morris, H.R., Dell, A. and Eichler, J., 2008. Identification of AglE, a second glycosyltransferase involved in N glycosylation of the Haloferax volcanii S-layer glycoprotein. Journal of bacteriology, 190(9), pp.3140-3146. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Yurist‐Doutsch, S., Abu‐Qarn, M., Battaglia, F., Morris, H.R., Hitchen, P.G., Dell, A. and Eichler, J., 2008. aglF, aglG and aglI, novel members of a gene island involved in the N‐glycosylation of the Haloferax volcanii S‐layer glycoprotein. Molecular microbiology, 69(5), pp.1234-1245. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Plavner, N. and Eichler, J., 2008. Defining the topology of the N-glycosylation pathway in the halophilic archaeon Haloferax volcanii. Journal of Bacteriology, 190(24), pp.8045-8052. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Abu-Qarn, M., Yurist-Doutsch, S., Giordano, A., Trauner, A., Morris, H.R., Hitchen, P., Medalia, O., Dell, A. and Eichler, J., 2007. Haloferax volcanii AglB and AglD are involved in N-glycosylation of the S-layer glycoprotein and proper assembly of the surface layer. Journal of molecular biology, 374(5), pp.1224-1236. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Abu‐Qarn, M. and Eichler, J., 2006. Protein N‐glycosylation in Archaea: defining Haloferax volcanii genes involved in S‐layer glycoprotein glycosylation. Molecular microbiology, 61(2), pp.511-525. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Eichler, J., 2000. Novel glycoproteins of the halophilic archaeon Haloferax volcanii. Archives of microbiology, 173(5), pp.445-448. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Mengele, R. and Sumper, M., 1992. Drastic differences in glycosylation of related S-layer glycoproteins from moderate and extreme halophiles. Journal of Biological Chemistry, 267(12), pp.8182-8185. |
| Corresponding Author | Jerry Eichler |
| Contact | Department of Life Sciences, Ben Gurion University of the Negev, PO Box 653, Beersheva 84105, Israel. |
| Reference | Sumper, M., Berg, E.L.K.E., Mengele, R.E.I.N.E.R. and Strobel, I.S.O.L.D.E., 1990. Primary structure and glycosylation of the S-layer protein of Haloferax volcanii. Journal of Bacteriology, 172(12), pp.7111-7118. |
