ProGlyProt ID AC108 Organism Information Organism Name Haloferax volcanii DS2 Domain Archaea Classification Family: Halobacteriaceae Order: Halobacteriales Class: Halobacteria or Halomebacteria Division or phylum: "Euryarchaeota" Taxonomic ID (NCBI) 2246 Genome Sequence (s) GeneBank CP001956.1 EMBL M62816 Gene Information Gene Name csg NCBI Gene ID 8924493 GenBank Gene Sequence 8924493 Protein Information Protein Name S-lyer glycoprotein UniProtKB/SwissProt ID P25062 NCBI RefSeq YP_003536097.1 EMBL-CDS AAA72996.1 UniProtKB Sequence >sp|P25062|CSG_HALVO Cell surface glycoprotein OS=Halobacterium volcanii GN=csg PE=1 SV=1 MTKLKDQTRAILLATLMVTSVFAGAIAFTGSAAAERGNLDADSESFNKTIQSGDRVFLGE EISTDAGLGASNPLLTGTAGNSEGVSLDLSSPIPQTTENQPLGTYDVDGSGSATTPNVTL LAPRITDSEILTSSGGDVTGSAISSSDAGNLYVNADYNYESAEKVEVTVEDPSGTDITNE VLSGTDTFVDDGSIGSTSSTGGGVGIDMSDQDAGEYTIILEGAEDLDFGDATETMTLTIS SQDEIGIELDSESVTQGTDVQYTVTNGIDGNEHVVAMDLSDLQNDATTEQAKEVFRNIGD TSEVGIANSSATNTSGSSTGPTVETADIAYAVVEIDGASAVGGIETQYLDDSEVDLEVYD AGVSATAAVGQDATNDITLTIEEGGTTLSSPTGQYVVGSEVDINGTATSSDSVAIYVRDD GDWQLLEIGGDNEISVDSDDTFEEEDIALSGLSGDGSSILSLTGTYRIGVIDASDADVGG DGSVDDSLTTSEFTSGVSSSNSIRVTDQALTGQFTTINGQVAPVETGTVDINGTASGANS VLVIFVDERGNVNYQEVSVDSDGTYDEDDITVGLTQGRVTAHILSVGRDSAIGDGSLPSG PSNGATLNDLTGYLDTLDQNNNNGEQINELIASETVDETASDDLIVTETFRLAESSTSID SIYPDAAEAAGINPVATGETMVIAGSTNLKPDDNTISIEVTNEDGTSVALEDTDEWNNDG QWMVEIDTTDFETGTFTVEADDGDNTDTVNVEVVSEREDTTTSSDNATDTTTTTDGPTET TTTAEPTETTEEPTEETTTSSNTPGFGIAVALVALVGAALLALRREN Sequence length 827 AA Subcellular Location Surface Function In Archaea, which do not possess other cell wall components, the S-layer has to maintain the cell integrity and stabilize as well as to protect the cell against mechanical and osmotic stresses or extreme pH conditions. It is also predicted that the S-layer has to maintain or even determine the cell shape. Protein Structure PDB ID Glycosylation Status Glycosylation Type N (Asn) linked, (O, Thr-linked residues not known) Experimentally Validated Glycosite(s) in Full Length Protein (Signal peptide: 1-34) N47, N117, N308, N313, N532 Experimentally Validated Glycosite(s ) in Mature Protein N13, N83, N274, N279, N498 Glycosite(s) Annotated Protein Sequence >sp|P25062|CSG_HALVO Cell surface glycoprotein OS=Halobacterium volcanii GN=csg PE=1 SV=1 MTKLKDQTRAILLATLMVTSVFAGAIAFTGSAAAERGNLDADSESFN*(47)KTIQSGDRVFLGE EISTDAGLGASNPLLTGTAGNSEGVSLDLSSPIPQTTENQPLGTYDVDGSGSATTPN*(117)VTL LAPRITDSEILTSSGGDVTGSAISSSDAGNLYVNADYNYESAEKVEVTVEDPSGTDITNE VLSGTDTFVDDGSIGSTSSTGGGVGIDMSDQDAGEYTIILEGAEDLDFGDATETMTLTIS SQDEIGIELDSESVTQGTDVQYTVTNGIDGNEHVVAMDLSDLQNDATTEQAKEVFRNIGD TSEVGIAN*(308)SSATN*(313)TSGSSTGPTVETADIAYAVVEIDGASAVGGIETQYLDDSEVDLEVYD AGVSATAAVGQDATNDITLTIEEGGTTLSSPTGQYVVGSEVDINGTATSSDSVAIYVRDD GDWQLLEIGGDNEISVDSDDTFEEEDIALSGLSGDGSSILSLTGTYRIGVIDASDADVGG DGSVDDSLTTSEFTSGVSSSNSIRVTDQALTGQFTTINGQVAPVETGTVDIN*(532)GTASGANS VLVIFVDERGNVNYQEVSVDSDGTYDEDDITVGLTQGRVTAHILSVGRDSAIGDGSLPSG PSNGATLNDLTGYLDTLDQNNNNGEQINELIASETVDETASDDLIVTETFRLAESSTSID SIYPDAAEAAGINPVATGETMVIAGSTNLKPDDNTISIEVTNEDGTSVALEDTDEWNNDG QWMVEIDTTDFETGTFTVEADDGDNTDTVNVEVVSEREDTTTSSDNATDTTTTTDGPTET TTTAEPTETTEEPTEETTTSSNTPGFGIAVALVALVGAALLALRREN Sequence Around Glycosites (21 AA) GNLDADSESFNKTIQSGDRVF VDGSGSATTPNVTLLAPRITD IGDTSEVGIANSSATNTSGSS EVGIANSSATNTSGSSTGPTV APVETGTVDINGTASGANSVL Glycosite Sequence Logo seqlogo Glycosite Sequence Logo 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 Linkage: βGlc-Asn, Gal-Thr. N glycosylated with a Pentasaccharide (2 hexoses, 2 hexuronic acids and a methyl ester of hexuronic acid). Pentasaccharide sequence is: hexose (Hex)-X-hexuronic acid (HexA)-HexA-Hex-peptide, where X is a 190 Da moiety, likely to be either dimethylated hexose or the methyl ester of hexuronic acid. Relatively minor amount of the completed pentasaccharide as compared to the level of tetrasaccharide as well as Di- and trisaccharide-bearing species were detected duri Technique(s) used for Glycan Identification Protein Glycosylation linked (PGL) gene(s) OST Gene Name aglB OST NCBI Gene ID 8923906 OST GenBank Gene Sequence 8923906 OST Protein Name AglB OST UniProtKB/ SwissProt ID D4GYH4 OST NCBI RefSeq YP_003535577.1 OST EMBL-CDS ADE03209.1 OST UniProtKB Sequence >tr|D4GYH4|D4GYH4_HALVD Oligosaccharyltransferase AglB OS=Haloferax volcanii (strain ATCC 29605 / DSM 3757 / IFO 14742 / NCIMB 2012 / DS2) GN=aglB PE=4 SV=1 MSDEQTKYSPSIAELARDWYHIPVLSTIILVMLWIRLRSYDAFIREGTVFFSGNDAWYHL RQVEYTVRNWPATMPFDPWTEFPFGRTAGQFGTIYDQLVATAALVVGLGSPSSDLVAKSL LVAPAVFGALTVIPTYLIGKRLGGRLGGLFGAVILMLLPGTFLQRGLVGFADHNIVEPFF MGFAVLAIMIALTVADREKPVWELVAARDLDALREPLKWSVLAGVATAIYMWSWPPGILL VGIFGLFLVLKMASDYVRGRSPEHTAFVGAISMTVTGLLMFIPIEEPGFGVTDFGFLQPL FSLGVALGAVFLAALARWWESNDVDERYYPAVVGGTMLVGIVLFSLVLPSVFDSIARNFL RTVGFSAGAATRTISEAQPFLAANVLQSNGQTAVGRIMSEYGFTFFTGALAAVWLVAKPL VKGGNSRKIGYAVGSLALIGVLFLIPALPAGIGSALGVEPSLVSLTIVTALIVGAVMQAD YESERLFVLVWAAIITSAAFTQVRFNYYLAVVVAVMNAYLLREALGIDFVGLANVERFDD ISYGQVAAVVIAVLLILTPVLIIPIQLGNGGVSQTAMQASQTGPGTVTQWDGSLTWMQNN TPAEGEFGGESNRMEYYGTYEYTDDFDYPDGAYGVMSWWDYGHWITVLGERIPNANPFQG GATEAANYLLAEDEQQAESVLTSMGDDGEGDQTRYVMVDWQMASTDAKFSAPTVFYDESN ISRSDFYNPMFRLQEQGEQTTVAAASSLKDQRYYESLMVRLYAYHGSAREASPIVVDWEE RTSADGSTTFRVTPSDGQAVRTFDNMSAAEEYVANDPTSQIGGIGTFPEE OST EC Number (BRENDA) 2.4.1.119 OST Genome Context 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. aglB is also present in this island. Characterized by a combination of gene deletion, Mass spectroscopy and biochemical characterization PGL Additional Links CAZy Literatures Reference(s) 1) 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. J Bacteriol, 192, 5572-5579. [PubMed: 20802039] 2) 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, 315108. [PubM Additional Comments 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 Year of Identification 1990 Year of Validation 1992