ProGlyProt ID BC111 Organism Information Organism Name Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) Domain Bacteria Classification Family: Bacteroidaceae Order: "Bacteroidales" Class: "Bacteroidia" Division or phylum: "Bacteroidetes" Taxonomic ID (NCBI) 272559 Genome Sequence (s) GeneBank CR626927.1 EMBL CR626927 Gene Information Gene Name BF0994 NCBI Gene ID 3287312 GenBank Gene Sequence 3287312 Protein Information Protein Name Hypothetical protein UniProtKB/SwissProt ID Q5LGK6 NCBI RefSeq YP_210682.1 EMBL-CDS CAH06733.1 UniProtKB Sequence >tr|Q5LGK6|Q5LGK6_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF0994 PE=4 SV=1 MATLQNIRSKGPLLVIVIGLALFAFIAGDAWKVLQPHQSHDVGEVNGETLSAQDYQNMVE EYTEVIKFSSGMSSLNDEQTNQVKDEVWRSYVNNKLIEKEAKKLGITVSKAEIQSIINEG VNPLLQQTPFRNPQTGAFDKDMLKKFLADYSKMDKTKMPSQYVEYYEGMHKLWSFVEKTL IQSRLAEKYQALVTKALFSNPVEAQDAFDARVNQSDVLLAAVPYSSIVDSTITVKESELK DLYNKKKEQFKQYVETRNIKYIDVQVIASAEDRAAIQQEVTDYTNQLATANGDYTTFIRS TGSEYPYVDLYYTKKAFPSDVVARMDSASIGQVYGPYYNAGDNTINSFKVLSKVAAADSV QFRQIQVYTEDAAKTKALADSIYTAIKGGADFTALAKKYGQTGESNWISSANYENAQVDG DNLKFISTINNLGVNELSNVALGQGNIILQVTDKKAVKDKYKVAVIKRAVEFSKETYNKA YNEFSQFIAANPTVDKVAANAEESGYKLLERNDLYSSEHGIGGIRGTKEALKWAFAAKPG EVSGLYECGESDRMLVVGLVSVIEEGYRPLAQVQDQLRAEIIRDKKAEKIMADMKAANAT TIAQYTSMANAVSDSVKHVTFAAPAYVAALRSSEPLVGAYASVSDINKLSAPIKGNGGVF VLQVYAKDKLNETFDAQSEEATLENMHARLASRFMNDLYLKGDVKDKRYLFF Sequence length 712 AA Subcellular Location Periplasm Function Predicted to function as a proline cis-trans isomerase (parvulin family) involved in a chaperone system. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Ser) linked Experimentally Validated Glycosite(s) in Full Length Protein S230, S327, S615 Experimentally Validated Glycosite(s ) in Mature Protein S230, S327, S615 Glycosite(s) Annotated Protein Sequence >tr|Q5LGK6|Q5LGK6_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF0994 PE=4 SV=1 MATLQNIRSKGPLLVIVIGLALFAFIAGDAWKVLQPHQSHDVGEVNGETLSAQDYQNMVE EYTEVIKFSSGMSSLNDEQTNQVKDEVWRSYVNNKLIEKEAKKLGITVSKAEIQSIINEG VNPLLQQTPFRNPQTGAFDKDMLKKFLADYSKMDKTKMPSQYVEYYEGMHKLWSFVEKTL IQSRLAEKYQALVTKALFSNPVEAQDAFDARVNQSDVLLAAVPYSSIVDS*(230)TITVKESELK DLYNKKKEQFKQYVETRNIKYIDVQVIASAEDRAAIQQEVTDYTNQLATANGDYTTFIRS TGSEYPYVDLYYTKKAFPSDVVARMDS*(327)ASIGQVYGPYYNAGDNTINSFKVLSKVAAADSV QFRQIQVYTEDAAKTKALADSIYTAIKGGADFTALAKKYGQTGESNWISSANYENAQVDG DNLKFISTINNLGVNELSNVALGQGNIILQVTDKKAVKDKYKVAVIKRAVEFSKETYNKA YNEFSQFIAANPTVDKVAANAEESGYKLLERNDLYSSEHGIGGIRGTKEALKWAFAAKPG EVSGLYECGESDRMLVVGLVSVIEEGYRPLAQVQDQLRAEIIRDKKAEKIMADMKAANAT TIAQYTSMANAVSDS*(615)VKHVTFAAPAYVAALRSSEPLVGAYASVSDINKLSAPIKGNGGVF VLQVYAKDKLNETFDAQSEEATLENMHARLASRFMNDLYLKGDVKDKRYLFF Sequence Around Glycosites (21 AA) AAVPYSSIVDSTITVKESELK FPSDVVARMDSASIGQVYGPY YTSMANAVSDSVKHVTFAAPA Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Mass shift detected on SDS-polyacrylamide gel, AAL (Aleuria aurantia lectin) reactivity, Pro-Q Emerald Glycostaining and reactivity towards anti-glycan antiserum Technique(s) used for Glycosylated Residue(s) Detection Site-directed mutagenesis Protein Glycosylation- Implication Protein glycosylation is central to the physiology of B. fragilis and is necessary for the organism to competitively colonize the mammalian intestine. Deletion of the lfg (protein glycosylation machinery) region results in a substantial growth deficiency in vitro and a complete inability to compete with wild-type bacteria in the mouse intestine. Glycan Information Glycan Annotation Exogenous fucose. Technique(s) used for Glycan Identification Lectin (AAL)binding Protein Glycosylation linked (PGL) gene(s) OST Gene Name wcfB OST NCBI Gene ID 3287270 OST GenBank Gene Sequence 3287270 OST Protein Name Putative fucosyl transferase OST UniProtKB/ SwissProt ID Q5LGI6 OST NCBI RefSeq YP_210702.1 OST EMBL-CDS CAH06753.1 OST UniProtKB Sequence >tr|Q5LGI6|Q5LGI6_BACFN Putative fucosyl transferase OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=wcfB PE=4 SV=1 MLYVILRGRLGNNLFQIATAASLTQNFIFCTVNKDQERQVLLYKDSFFKNIKVMKGVPDG IPYYKEPFHEFSRIPYEEGKDLIIDGYFQSEKYFKRSVVLDLYRITDELRKKIWNICGNI LEKGETVSIHVRRGDYLKLPHALPFCGKSYYKNAIQYIGEDKIFIICSDDIDWCKKNFIG KRYYFIENTTPLLDLYIQSLCTHNIISNSSFSWWGAWLNENSNKIVIAPQMWFGISVKLG VSDLLPVSWVRLPNNYTLGRYCFALYKVVEDYLLNILRLIWKRKKNM OST EC Number (BRENDA) OST Genome Context Q5LGI6 Characterized Accessory Gene(s) PGL Additional Links CAZy Literatures Reference(s) 1) Fletcher, C.M., Coyne, M.J. and Comstock, L.E. (2011) Theoretical and experimental characterization of the scope of protein O-glycosylation in Bacteroides fragilis. J Biol Chem, 286, 3219-3226. [PubMed: 21115495] 2) Fletcher, C.M., Coyne, M.J., Villa, O.F., Chatzidaki-Livanis, M. and Comstock, L.E. (2009) A general O-glycosylation system important to the physiology of a major human intestinal symbiont. Cell, 137, 321-331. [PubMed: 19379697] Additional Comments Glycosylation sequon features: the sequon has an aspartate (D) preceding the glycosylated T or S which is followed by an amino acid with one or more methyl groups (alanine, isoleucine, or leucine; (D)(S/T)(A/I/L/V/M/T). Moreover, none of the 17 unglycosylated S and T residues examined in of BF2494 (excluding two in the signal peptide) have a preceding D, although seven are followed by A, I, or L and one by V. Non methylated amino acids were not tolerated at third position of sequon in BF2494. Il Year of Identification 2009 Year of Validation 2011