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ProGlyProt ID
AC120
Organism Information
Organism Name
Sulfolobus acidocaldarius (DSM 639)
Domain
Archaea
Classification
Family: Sulfolobaceae
Order: Sulfolobales
Class: Thermoprotei or Crenarchaeota
Division or phylum: "Crenarchaeota"
Taxonomic ID (NCBI)
 
 
Genome Sequence(s)
GenBank
EMBL
 
 
Gene Information
Gene Name
cbsA(Saci_1858)
NCBI Gene ID
GenBank Gene Sequence
 
 
Protein Information
Protein Name
Cytochrome b558/566 subunit A (b type hemoprotein)
UniProtKB/SwissProt ID
NCBI RefSeq
EMBL-CDS
UniProtKB Sequence
Sequence length
462 AA
Subcellular Location
Integral 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 Ser/Thr- linked residues not yet identified)
Experimentally Validated Glycosite(s) in Full Length Protein
N144, N164
Experimentally Validated Glycosite(s ) in Mature Protein
N144, N164
Glycosite(s) Annotated Protein Sequence
>sp|O54088|CBSA_SULAC Cytochrome b558/566 subunit A OS=Sulfolobus acidocaldarius
GN=cbsA PE=1 SV=1 MSLKIKSKITIGVLLIIFLLSIIFTLENVSLAQTSPQISVYKVVGSADLSNPGSAG
YWSQ IPWTNISLTANIPMAPTSGLTHYLLVKAAWNGSWIFILEEWQAPEPAFNAWSTAVAGIYP NASGPGLF
RMIELTPGTTYSLERN*(144)YTNYVSIINGKEETGRIVFN*(164)YSGITLPAPNNTEITV MSNGTI
LLWHSPRPVEDLLYNDGMFYGYYVNSTWYYPDRAAIMWYLGSGVPTKDDMNIGG KYPGQQFDGITFKDAGGSLA
QSGGSANIWMWVSGATWNNSTYDPAFKSNIWQNESLTGLS YVDSGNHGFAVPLYTNNTNMYEVDTAGIWYTPVA
SEGLNGSLFFIWTGAKYENGSWVVEF ARPLSVPLDYQPFMPNITVGKTYYVAFAVWQGRLGETLFDKSITSSFL
SLELVTTPPTST TTSTSPVTTISSAIPPVTLYVTIIGVVVALVALVILYVVFRR
Sequence Around Glycosites (21 AA)
TPGTTYSLERNYTNYVSIING
GKEETGRIVFNYSGITLPAPN
Glycosite Sequence Logo
Glycosite Sequence Logo
Technique(s) used for Glycosylation Detection
Chemical 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 ID
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 Name
Saci_1274
OST NCBI Gene ID
OST GenBank Gene Sequence
OST Protein Name
Conserved membrane protein
OST UniProtKB/ SwissProt ID
OST NCBI RefSeq
OST EMBL-CDS
OST UniProtKB Sequence
>tr|Q4J9B4|Q4J9B4_SULAC Conserved membrane protein OS=Sulfolobus acidocaldarius
GN=Saci_1274 PE=4 SV=1 MQSTSILARIDKFKFLDAIIIGSLALFSILIRIISITAFPQTINGFDSWYLFY
NALLIVK AGGNWYAVPPDVHAWFPWGYFIELENTIGLPFLVALFSVPFYSIFGQNIVYTLTLVSPIV LDGIGVV
AAFLAVESITNSRVGGYIAAAITAFTPSLTYKNILGSLPKTSWGGVFVLFTIY FLSLAIKKKKPLYGIPAGIMIF
LANITWGGYTYIDISLAIAAFLIVLFNKNDEISAKTLT ISGITAAFLTSLSPNTIGFMSEVAHGLALLIIPLFLY
LDLYLRRVLPKDIVDSKNIVIGA AIILLVSLVVLGSVAFKVQLIPSRYYAIINPFFQFTVPIDRTVAEYIPQSIA
AMIQDFGI GLFLSIIGIYFLLTRKQDMAGIWLVVLGAASIYGTSEQPYLFNYTIYIVAALAGVAVAEL FSRFME
RKIRIAPILMLTLIGVALLADAGIAVEASYAPQALINSSTSYLTTNYAWISALD WINQNTPNNAFILSWWDYGYW
IHAVGNRTVIDENNTLNGTQIKLMAEMFLNNESFAVNVL ENDFHLYPYGNPNYTRPVYIVAYDAVTEYIVNNQYP
VWFIGYPTNFPGTFIGYTTSLGDI AKAIGAMTTIAGYNTNSYVNTTYINETASYAAQYNSQLASIIANSLPMAWT
PKTYNSLIG SMFIEAIQSLNQGPVQAPFSISLSQLLQSSSTSLYNPNALLPRVNLMYFKPVYIALFPLS VTNAL
GGEAIVYIMVYIYQFVMPNVIIPPTISTA
OST EC Number (BRENDA)
OST Genome Context
PGL Additional Links
 
 
Literature
Reference(s)
1) 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]
2) 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]
Additional Comments
Provides 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 Identification
1998
Year of Validation
2000
 
 
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