ProGlyProt ID BC101 Organism Information Organism Name Acetogenium kivui (Thermoanaerobacter kivui) Domain Bacteria Classification Family: Thermoanaerobacteraceae Order: Thermoanaerobacterales Class: Clostridia Division or phylum: "Firmicutes" Taxonomic ID (NCBI) 2325 Genome Sequence (s) EMBL M31069 Gene Information Gene Name NCBI Gene ID Protein Information Protein Name Cell surface protein/ S-layer protein UniProtKB/SwissProt ID P22258 NCBI RefSeq EMBL-CDS AAA21930.1 UniProtKB Sequence >sp|P22258|SLAP_ACEKI Cell surface protein OS=Acetogenium kivui PE=1 SV=1 MKNLKKLIAVVSTFALVFSAMAVGFAATTPFTDVKDDAPYASAVARLYALNITNGVGDPK FGVDQPVTRAQMITFVNRMLGYEDLAEMAKSEKSAFKDVPQNHWAVGQINLAYKLGLAQG VGNGKFDPNSELRYAQALAFVLRALGFKDLDWPYGYLAKAQDLGLVHGLNLAYNGVIKRG DLALILDRALEVPMVKYVDGKEVLGEPLISKVATKAEYTVIATNAQDRSVEEGKVAVLDK DGKLTTINAGLVDFSEYLGKKVIVYSERFGDPVYVAEGDNDVVSFTEGQDSVGTTVYKND DNKTAIKVDDNAYVLYNGYLTKVSKVTVKEGAEVTIINNNYLIVNGSYDNSTIVYNDVQS GDKYLNRDSNYELKGTVTVTGAVSKVTDIKANDYIYYGKQYDVNGNVVGTVIYVVRNQVT GTVTEKSVSGSTYKASIDNVSYTVADNNVWNQLEPGKKVTVILNKDNVIVGISSTTTTTA VNYAIFKEKSDPFTAWFAKVKLILPDAAEKVFDAVYSDVYDKVNLAEGTIVTYTVDANGK LNDIQRANDQPFSSASYKADAKVLTEGSTTYYITDNTVLLNNTSDGYKALKLTDLKDATN LNVKIVADNYNVAKVVVFNNASFVSTTTSTVYAYVTGTADVYVNGSTFTRLTVLENGQTK TYDANAQLATNYTHKAVVLTLTNAKIANIALPTVASGVKLTNIDQANLRITDTTNKGYLL DPNFIVVDTNGNLKGLSDITKDTGVNLYTNDVGKVFVIEIVQ Sequence length 762 AA Subcellular Location Surface Function The S-layer is a paracrystalline mono-layered assembly of proteins which coat the surface of bacteria. This bacterium is covered by a S-layer with hexagonal symmetry. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Tyr) linked Experimentally Validated Glycosite(s) in Full Length Protein Y297, Y516, Y520, Y632 Experimentally Validated Glycosite(s ) in Mature Protein Y297, Y516, Y520, Y632 Glycosite(s) Annotated Protein Sequence >sp|P22258|SLAP_ACEKI Cell surface protein OS=Acetogenium kivui PE=1 SV=1 MKNLKKLIAVVSTFALVFSAMAVGFAATTPFTDVKDDAPYASAVARLYALNITNGVGDPK FGVDQPVTRAQMITFVNRMLGYEDLAEMAKSEKSAFKDVPQNHWAVGQINLAYKLGLAQG VGNGKFDPNSELRYAQALAFVLRALGFKDLDWPYGYLAKAQDLGLVHGLNLAYNGVIKRG DLALILDRALEVPMVKYVDGKEVLGEPLISKVATKAEYTVIATNAQDRSVEEGKVAVLDK DGKLTTINAGLVDFSEYLGKKVIVYSERFGDPVYVAEGDNDVVSFTEGQDSVGTTVY*(297)KND DNKTAIKVDDNAYVLYNGYLTKVSKVTVKEGAEVTIINNNYLIVNGSYDNSTIVYNDVQS GDKYLNRDSNYELKGTVTVTGAVSKVTDIKANDYIYYGKQYDVNGNVVGTVIYVVRNQVT GTVTEKSVSGSTYKASIDNVSYTVADNNVWNQLEPGKKVTVILNKDNVIVGISSTTTTTA VNYAIFKEKSDPFTAWFAKVKLILPDAAEKVFDAVY*(516)SDVY*(520)DKVNLAEGTIVTYTVDANGK LNDIQRANDQPFSSASYKADAKVLTEGSTTYYITDNTVLLNNTSDGYKALKLTDLKDATN LNVKIVADNYNVAKVVVFNNASFVSTTTSTVY*(632)AYVTGTADVYVNGSTFTRLTVLENGQTK TYDANAQLATNYTHKAVVLTLTNAKIANIALPTVASGVKLTNIDQANLRITDTTNKGYLL DPNFIVVDTNGNLKGLSDITKDTGVNLYTNDVGKVFVIEIVQ Sequence Around Glycosites (21 AA) EGQDSVGTTVYKNDDNKTAIK DAAEKVFDAVYSDVYDKVNLA KVFDAVYSDVYDKVNLAEGTI SFVSTTTSTVYAYVTGTADVY Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Deglycosylation (trifluoromethanesulfonic acid), aberrant migration on SDS-PAGE. Technique(s) used for Glycosylated Residue(s) Detection Protease digestion and glycopeptide separation by molecular sieve chromatography and reversed-phase HPLC. Deglycosylated and undeglycosylated peptides were run on 2D PAGE and subjected to N-terminal sequencing post blotting. Protein Glycosylation- Implication An N terminal SLH sequence of A. kivui S layer glycoprotein is suggested to fold independently and present itself in a modular way that may serve as an anchor to the peptidoglycan. Glycosylated residues are surface exposed and implicated in protease K resistance. Glycan Information Glycan Annotation Carbohydrate content 8% corresponding to 40 to 50 sugar molecules per protein monomer. Acidic, repetitive heterogenous glycans of varying length containing N-acetylglucosamine, 1,6 linked glucose, 1,4 linked galactosamine and/or unidentified sugar-related component. Technique(s) used for Glycan Identification Hydrazinolysis followed by amino-acid analysis as well as two-dimensional NMR spectroscopy Protein Glycosylation linked (PGL) gene(s) OST Gene Name OST NCBI Gene ID OST GenBank Gene Sequence OST Protein Name OST UniProtKB/ SwissProt ID OST NCBI RefSeq OST EMBL-CDS OST UniProtKB Sequence OST EC Number (BRENDA) OST Genome Context Characterized Accessory Gene(s) PGL Additional Links CAZy Literatures Reference(s) 1) Lupas, A., Engelhardt, H., Peters, J., Santarius, U., Volker, S. and Baumeister, W. (1994) Domain structure of the Acetogenium kivui surface layer revealed by electron crystallography and sequence analysis. J Bacteriol, 176, 1224-1233. [PubMed: 8113161] 2) Peters, J., Rudolf, S., Oschkinat, H., Mengele, R., Sumper, M., Kellermann, J., Lottspeich, F. and Baumeister, W. (1992) Evidence for tyrosine-linked glycosaminoglycan in a bacterial surface protein. Biol Chem Hoppe Seyler, 373, 171-17 Additional Comments A new recognition sequence for tyrosine linked glycosylation in A. kivui is suggested based on the fact that all glycosylated tyrosine in the sequence are preceded by valine. Year of Identification 1989 Year of Validation 1992