ProGlyProt ID AC102 Organism Information Organism Name Halobacterium salinarum (halobium) R1M1/NRC-1 Domain Archaea Classification Family: Halobacteriaceae Order: Halobacteriales Class: Halobacteria or Halomebacteria Division or phylum: "Euryarchaeota" Taxonomic ID (NCBI) 64091 Genome Sequence (s) GeneBank AE004437.1 EMBL AE004437 Gene Information Gene Name csg (VNG_2679G) NCBI Gene ID 1449008 GenBank Gene Sequence 1449008 Protein Information Protein Name S layer glycoprotein UniProtKB/SwissProt ID Q9HM69 NCBI RefSeq NP_281222.1 EMBL-CDS AAG20702.1 UniProtKB Sequence >sp|Q9HM69|CSG_HALSA Cell surface glycoprotein OS=Halobacterium salinarium GN=csg PE=1 SV=2 MTDTTGKLRAVLLTALMVGSVIGAGVAFTGGAAAANASDLNDYQRFNENTNYTYSTASED GKTEGSVASGATIFQGEEDVTFRKLDNEKEVSPATLSRTGGSDEGVPLQMPIPEDQSTGS YDSNGPDNDEADFGVTVQSPSVTMLEVRNNADNDVTGGVLNTQQDESSIAVDYNYYAAED LELTVEDEDGLDVTDEILAADQSGGAYEDGTGNNGPNTLRFDIDPNNVDAGDYTVSVEGV EDLDFGDATESASVTISSSNKASLNLAEDEVVQGANLKYTIENSPEGNYHAVTIDSSDFR DSSSGADAAKVMRSVGDTVDTGLVVDNDSTTEIVDDYENTSISDVDYAYAIVEIDDGNGV GSIETQYLDDSSADIDLYPASDTEDAPDYVNSNEELTNGSALDGVSTDDDTDFDVTQGDI TLDNPTGAYVVGSEVDINGTANEGTDDVVLYARDNNDFELVTVDGEKSIEVDSDDTFEEE DITLSDGDKGGDDILGLPGTYRLGIIAKSDAVNSSGGVKDNIDTSDFNQGVSSTSSIRVT DTELTASFETYNGQVADDDNQIDVEGTAPGKDNVAAIIIGSRGKVKFQSISVDSDDTFDE EDIDISELRQGSASAHILSSGRDGKFGEDTANSISDLEDEVGNYTSGSPTGDQIRDRILS NTVDDTASDDLIVTQQFRLVDGLTTIEATEGGEAGGSLTVMGTTNRKADDNTITVELLQG DASIEINSTDEWNSDGQWSVDVPLSNVEPGNYTVEADDGDNTDRQNVEIVEELEEPDQTT VDQPENNQTMTTTMTETTTETTTEMTTTQENTTENGSEGTSDGESGGSIPGFGVGVALVA VLGAALLALRQN Sequence length 852 AA Subcellular Location Surface Function In Archaea, which do not possess other cell wall components, the S-layer has been implicated in the maintaineance of 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) N36, N339, N398, N438, N513, N643, N727, N751, N787, N811, N815 (N36, N513 and N643 were confirmed glycosylated directly by glycopeptide sequence analysis , the reference no. 2 mentions that ten sulfated saccharides are N-linked to the protein imlplying that most or all sites are glycosylated.) Experimentally Validated Glycosite(s ) in Mature Protein N2, N305, N364, N404, N479, N609, N693, N717, N753, N777, N781 Glycosite(s) Annotated Protein Sequence >sp|Q9HM69|CSG_HALSA Cell surface glycoprotein OS=Halobacterium salinarium GN=csg PE=1 SV=2 MTDTTGKLRAVLLTALMVGSVIGAGVAFTGGAAAAN*(36)ASDLNDYQRFNENTNYTYSTASED GKTEGSVASGATIFQGEEDVTFRKLDNEKEVSPATLSRTGGSDEGVPLQMPIPEDQSTGS YDSNGPDNDEADFGVTVQSPSVTMLEVRNNADNDVTGGVLNTQQDESSIAVDYNYYAAED LELTVEDEDGLDVTDEILAADQSGGAYEDGTGNNGPNTLRFDIDPNNVDAGDYTVSVEGV EDLDFGDATESASVTISSSNKASLNLAEDEVVQGANLKYTIENSPEGNYHAVTIDSSDFR DSSSGADAAKVMRSVGDTVDTGLVVDNDSTTEIVDDYEN*(339)TSISDVDYAYAIVEIDDGNGV GSIETQYLDDSSADIDLYPASDTEDAPDYVNSNEELTN*(398)GSALDGVSTDDDTDFDVTQGDI TLDNPTGAYVVGSEVDIN*(438)GTANEGTDDVVLYARDNNDFELVTVDGEKSIEVDSDDTFEEE DITLSDGDKGGDDILGLPGTYRLGIIAKSDAVN*(513)SSGGVKDNIDTSDFNQGVSSTSSIRVT DTELTASFETYNGQVADDDNQIDVEGTAPGKDNVAAIIIGSRGKVKFQSISVDSDDTFDE EDIDISELRQGSASAHILSSGRDGKFGEDTANSISDLEDEVGN*(643) YTSGSPTGDQIRDRILS NTVDDTASDDLIVTQQFRLVDGLTTIEATEGGEAGGSLTVMGTTNRKADDNTITVELLQG DASIEIN*(727)STDEWNSDGQWSVDVPLSNVEPGN*(751)YTVEADDGDNTDRQNVEIVEELEEPDQTT VDQPENN*(787)QTMTTTMTETTTETTTEMTTTQEN*(811)TTEN*(815)GSEGTSDGESGGSIPGFGVGVALVA VLGAALLALRQN Sequence Around Glycosites (21 AA) VAFTGGAAAANASDLNDYQRF STTEIVDDYENTSISDVDYAY DYVNSNEELTNGSALDGVSTD AYVVGSEVDINGTANEGTDDV LGIIAKSDAVNSSGGVKDNID SISDLEDEVGNYTSGSPTGDQ LLQGDASIEINSTDEWNSDGQ DVPLSNVEPGNYTVEADDGDN DQTTVDQPENNQTMTTTMTET TTTEMTTTQENTTENGSEGTS MTTTQENTTENGSEGTSDGES Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Periodate-arsenite-Schiff reagent staining and carbohydrate analysis using GC. Technique(s) used for Glycosylated Residue(s) Detection Glycopeptide sequencing Protein Glycosylation- Implication It is the pattern and the chemical nature of the N-linked saccharides which exhibits a drastic change at the transition from moderate to extreme halophily. This different pattern of glycosylation (sulfated glucuronic acids and a repeating unit saccharide) introduces at least 120 additional negative charges into the glycoprotein. The protruding highly negatively charged loops are required for stabilization in high salt concentrations. Thus, the sulfated repeating unit saccharide is required for s Glycan Information Glycan Annotation Linkages: βGalNAc- Asn, Glc-Asn, Gal-Thr. Total carbohydrate content is approximately 10 to 12% of 200kDa S layer glycoprotein. N glycosylated (at position N2) with GlcNAc-linked repeating (10-15 repeats) sulfated pentasaccharide and with sulphated oligosaccharides: (GlcA(OSO3-)-[β1→4GlcA(OSO3-)]2-β1→4Glc-Asn) at ten other poistions. Abut 20 neutral di/ tri- saccharides α-D-Glc-(1→2)-Gal-(1→ or (uronic acid, glucose)-galactose are O-glycosidically attached to clustere Technique(s) used for Glycan Identification GC-MS (gas chromatography-mass spectrometry) analysis of peracetylated alditols. Protein Glycosylation linked (PGL) gene(s) OST Gene Name aglB/stt3 OST NCBI Gene ID 5953829 OST GenBank Gene Sequence 5953829 OST Protein Name AglB/STT3 subunit OST UniProtKB/ SwissProt ID B0R4T2 OST NCBI RefSeq YP_001689095.1 OST EMBL-CDS CAP13747.1 OST UniProtKB Sequence >tr|B0R4T2|B0R4T2_HALS3 Oligosaccharyl transferase OS=Halobacterium salinarum (strain ATCC 29341 / DSM 671 / R1) GN=stt3 PE=4 SV=1 MSETTDASGLSAGLPSRALGALKDWYHVPVLGAVLAFMFWVRVQSWDNFLQNGEVYLSGN DAYYHLRQVTYTVHHFPETMPFDPWTNFPHGTLAGQFGTLFDQILAAVALVVGMGSPSEH TIAMTLLLAPPVFGALLVVPTYVMGRRIAGRLGGLFGAVILGLLPGYFLQRTLAGAADHN GAEPLFMTLAVVGLMVALSVAEREKPVFEQFRAWDATGLRDVVGWSVLAGVATAVYMWVW PPGVLLVGIFGAFFLVKLVGDYVTGTSPDHVAVVGAVSMTTTALLMFAPLGESGFGVSGF SFLQPVFALGVAVGCVVLAWLARVFDERSLSPAAYVATVAGTLVVVVGALALVAPGFWSS LQSNLLNYIGLSSTADTRTIGEAQPFLFRTGRYGLGMFGVVFLEYGTAFFSALVGAGAIL LKPHLTSGSVRRIVGVAGATAVLAIVAVSPAVPAAIGGVVGLGGQLTALLIAAVVLAAIA ATGQYDADKLLLVVWGAFVTSAAFTQVRFNYYLVVPVVVLNAYVLRAVLAAVDLDRPVAA IEGVSWYQVGTVVIVALIVLVPVAAPAVAEATNNDSASGPVSPIKLSNNQNQPVPLQSAI TAGQGQGPGGVVAWDDAMDWYQSHTPKQGTYGGADNADALDYNGTYSQTEDFDYPAGSFG VLSWWDYGHWMTVQGHAIPHANPFQQGATSAANFLLADDEQQSESVLADIEEDDAKNRYV AVDWKMVMPPLPYLSSGQSKFSAPVVFYDGPRDLSQGDFYQQAYNADWEENRISNARFFR QSPLLKKQAYYESMMVRLYRYHGSAKQAEPVVADWRRSVNTGQGTAAAFDAATNNTKQFD SMADAKAYVRNDSTA OST EC Number (BRENDA) 2.4.1.119 OST Genome Context Characterized Accessory Gene(s) PGL Additional Links CAZy Literatures Reference(s) 1) Mengele, R. and Sumper, M. (1992) Drastic differences in glycosylation of related S-layer glycoproteins from moderate and extreme halophiles. J Biol Chem, 267, 8182-8185. [PubMed: 1569073] 2) Lechner, J. and Sumper, M. (1987) The primary structure of a procaryotic glycoprotein. Cloning and sequencing of the cell surface glycoprotein gene of halobacteria. J Biol Chem, 262, 9724-9729. [PubMed: 3036870] 3) Paul, G., Lottspeich, F. and Wieland, F. (1986) Asparaginyl-N-acetylgalactosamin Additional Comments First prokaryotic glycoprotein that was charcterized experimentally for the site of glycosylation as well as glycan attached. The features unique to N glycosylation in H salinarium are: the majority of glycans are linked via glucose instead of GlcNAc or GalNAc to the Asn in protein. Presence of sulfated oligosaccharides that bind to a C60-dolichol monophosphate carrier lipid. Protein associated glycans differ mainly in terminal sugars. S layer proteins of H salinarium and H. volcanii are the exa Year of Identification 1974 Year of Validation 1987