ProGP151

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ProGP ID ProGP151
Validation Status Characterized
Organism Information
Organism NameSulfolobus acidocaldarius (DSM 639)
Domain Archaea
Classification Family: Sulfolobaceae
Order: Sulfolobales
Class: Thermoprotei or Crenarchaeota
Division or phylum: "Crenarchaeota"
Taxonomic ID (NCBI) 2285
Genome Sequence(s)
GenBank CP000077.1
EMBL CP000077
Gene Information
Gene NamecbsA(Saci_1858)
NCBI Gene ID 3474781
GenBank Gene Sequence 3474781
Protein Information
Protein NameCytochrome b558/566 subunit A (b type hemoprotein)
UniProtKB/SwissProt ID O54088
NCBI RefSeq YP_256457.1
EMBL-CDSAAY81164.1
UniProtKB Sequence >sp|O54088|CBSA_SULAC Cytochrome b558/566 subunit A OS=Sulfolobus acidocaldarius GN=cbsA PE=1 SV=1 MSLKIKSKITIGVLLIIFLLSIIFTLENVSLAQTSPQISVYKVVGSADLSNPGSAGYWSQ IPWTNISLTANIPMAPTSGLTHYLLVKAAWNGSWIFILEEWQAPEPAFNAWSTAVAGIYP NASGPGLFRMIELTPGTTYSLERNYTNYVSIINGKEETGRIVFNYSGITLPAPNNTEITV MSNGTILLWHSPRPVEDLLYNDGMFYGYYVNSTWYYPDRAAIMWYLGSGVPTKDDMNIGG KYPGQQFDGITFKDAGGSLAQSGGSANIWMWVSGATWNNSTYDPAFKSNIWQNESLTGLS YVDSGNHGFAVPLYTNNTNMYEVDTAGIWYTPVASEGLNGSLFFIWTGAKYENGSWVVEF ARPLSVPLDYQPFMPNITVGKTYYVAFAVWQGRLGETLFDKSITSSFLSLELVTTPPTST TTSTSPVTTISSAIPPVTLYVTIIGVVVALVALVILYVVFRR
Sequence length 462 AA
Subcellular LocationIntegral membrane protein
Function Membrane residing ectoenzyme. Due to the likely pseudoperiplasmic location, cytochrome c-like function is proposed analogous to the situation in Paracoccus denitrificans linking pseudoperiplasmic redox metabolism to membrane-residing electron transport systems. Cytochrome b558/566 is significantly up-regulated under certain organotrophic growth conditions in combination with low oxygen tension.
Glycosylation Status
Glycosylation Type N (Asn) linked (O linked residues not yet identified)
Experimentally Validated Glycosite(s) in Full Length ProteinN144, N164
Experimentally Validated Glycosite(s ) in Mature ProteinN144, N164
Glycosite(s) Annotated Protein Sequence >sp|O54088|CBSA_SULAC Cytochrome b558/566 subunit A OS=Sulfolobus acidocaldarius GN=cbsA PE=1 SV=1 MSLKIKSKITIGVLLIIFLLSIIFTLENVSLAQTSPQISVYKVVGSADLSNPGSAGYWSQ IPWTNISLTANIPMAPTSGLTHYLLVKAAWNGSWIFILEEWQAPEPAFNAWSTAVAGIYP NASGPGLFRMIELTPGTTYSLERN*(144)YTNYVSIINGKEETGRIVFN*(164)YSGITLPAPNNTEITV MSNGTILLWHSPRPVEDLLYNDGMFYGYYVNSTWYYPDRAAIMWYLGSGVPTKDDMNIGG KYPGQQFDGITFKDAGGSLAQSGGSANIWMWVSGATWNNSTYDPAFKSNIWQNESLTGLS YVDSGNHGFAVPLYTNNTNMYEVDTAGIWYTPVASEGLNGSLFFIWTGAKYENGSWVVEF ARPLSVPLDYQPFMPNITVGKTYYVAFAVWQGRLGETLFDKSITSSFLSLELVTTPPTST TTSTSPVTTISSAIPPVTLYVTIIGVVVALVALVILYVVFRR
Sequence Around Glycosites (21 AA) TPGTTYSLERNYTNYVSIING
GKEETGRIVFNYSGITLPAPN
Glycosite Sequence Logo
Technique(s) used for Glycosylation DetectionChemical deglycosylation (with TFMS) and mass shift on SDS-PAGE.
Technique(s) used for Glycosylated Residue(s) Detection MALDI-MS, 1H-NMR spectroscopy of trypsin/pronase digested, anion-exchange HPLC purified glycan linked dipeptides.
Protein Glycosylation- Implication The increased glycosylation of Cytochrome b558/566 subunit A has been suggested to have a role in the protection of enzyme against the harsh acidic conditions (pH 2 - 2.5) of S. acidocaldarius, in its thermoacidophilic habitat. Upon glycosylation with a low pKa sulfonate carrying glycans the protein achieves a negative surface potential at the locus of the cytochrome. The glycosylation of a major internal loop region also may improve resistance against hydrolytic cleavage which is even better than what is acheived from sulfated glycans.
Glycan Information
Glycan Annotation Linkage: β-GlcNAc-Asn.
64-kDa glycoprotein expresses 17% glycosylation. Total sugar analysis revealed the relative proportions of approximately 7:2:2 for mannose, glucose and GlcNAc.
N-linked hexasaccharide units (at least seven)- two residues each of Man and GlcNAc (one trisubstituted) and one residue each of Glc and 6-deoxy-6-sulfoglucose (6-sulfoquinovose- a rare acidic sugar).
β-D-Glcp-(1→4)-β-D-Quip(6--SO3-)-(1→3)-[α-D-Manp(1→4), α-D-Manp(1→6)] β-D-GlcpNAc-(1→4)-β-D-GlcpNAc-(1→Asn....
O-mannosylation with atleast 35 monosaccharides at unidentified residues.
BCSDB ID5950
Technique(s) used for Glycan Identification Identification of oligosaccharides using Gel Chromatography and GLC-MS (PNGase F treatment followed by HPAEC analysis). Glycan structure is elucidated with the help of 1H-NMR spectrum from two-dimensional 1H, 1H correlation NMR Spectroscopy . Linkage analysis was carried out using 13C NMR Chemical Shift analysis.
Protein Glycosylation linked (PGL) gene(s)
OST Gene NameConserved membrane protein
OST ProGT IDProGT52
Accessory Gene(s)Progt IDProGT52.1-ProGT52.3
Literature
Additional CommentProvides first extensive study and a structure for oligosaccharide attached to a crenarchaeotal glycoprotein. The protein attached hexasaccharide is unique in terms of the presence of a trisubstituted N acetylglucosamine residue and a rare acidic residue Qui6S (sulfonated sugar).
Qui6S like sugar has previously been observed only in glycolipids of chloroplast and photosynthetic bacteria. Only other case where sufated sugar has been observed is H. salinaium.
Unusally, only one type of glycan has been found associated with cytochrome b558/566 subunit A.
Year of Identification1998
Year of Identification Month Wise1998.05.15
Year of Validation 2000
Reference Hettmann, T., Schmidt, C.L., Anemuller, S., Zahringer, U., Moll, H., Petersen, A. and Schafer, G. (1998) Cytochrome b558/566 from the archaeon Sulfolobus acidocaldarius. A novel highly glycosylated, membrane-bound b-type hemoprotein. J Biol Chem, 273, 12032-12040. [PubMed: 9575144]
Author Hettmann, T., Schmidt, C.L., Anemuller, S., Zahringer, U., Moll, H., Petersen, A. Schafer, G.
Research GroupInstitut für Biochemie, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Corresponding Author Schafer, G.
ContactInstitut für Biochemie, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Reference Hettmann, T., Schmidt, C.L., Anemuller, S., Zahringer, U., Moll, H., Petersen, A. and Schafer, G. (1998) Cytochrome b558/566 from the archaeon Sulfolobus acidocaldarius. A novel highly glycosylated, membrane-bound b-type hemoprotein. J Biol Chem, 273, 12032-12040. [PubMed: 9575144]
Author Hettmann, T., Schmidt, C.L., Anemuller, S., Zahringer, U., Moll, H., Petersen, A. and Schafer, G.
Research GroupInstitut für Biochemie, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Corresponding Author Schafer, G.
ContactInstitut für Biochemie, Medizinische Universität zu Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Reference1) Zahringer, U., Moll, H., Hettmann, T., Knirel, Y.A. and Schafer, G. (2000) Cytochrome b558/566 from the archaeon Sulfolobus acidocaldarius has a unique Asn-linked highly branched hexasaccharide chain containing 6-sulfoquinovose. Eur J Biochem, 267, 4144-4149. [PubMed: 10866817]
Author Zahringer, U., Moll, H., Hettmann, T., Knirel, Y.A. and Schafer, G.
Research GroupForschungszentrum Borstel, Zentrum für Medizin und Biowissenschaften, Borstel, Germany. uzaehr@fz-borstel.de
Corresponding Author Schafer, G.
ContactForschungszentrum Borstel, Zentrum für Medizin und Biowissenschaften, Borstel, Germany. uzaehr@fz-borstel.de