ProGlyProt ID BC110 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 BF0522 NCBI Gene ID 3286009 GenBank Gene Sequence 3286009 Protein Information Protein Name Hypothetical protein UniProtKB/SwissProt ID Q5LHV3 NCBI RefSeq YP_210235.1 EMBL-CDS CAH06277.1 UniProtKB Sequence >tr|Q5LHV3|Q5LHV3_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF0522 PE=4 SV=1 MKRLIPILLAVFAFAACEKDPDMDKLDNDYLVYTNYDKKADFKQFSTYYIPDSVLVIGDK KDPEYWKGEAAEAIINAYKENLNSKGFTYTDNKDAADLGIQVSYVQSTYYFTDYGQPEWW WNYPGYWDAPYWGNWGGWYYPYVVNYSITTNSFLTEIMNLKAPEGEKQKLPVLWSSFLSG PASYSGKVNQTLVVRAINQSFAQSPYLTNK Sequence length 210 AA Subcellular Location Outer membrane (facing surface) Function It is suggested to be involved in the response to oxidative stress as it was found upregulated 6.1-fold on exposure of B. fragilis to air. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Ser) linked Experimentally Validated Glycosite(s) in Full Length Protein S53 Experimentally Validated Glycosite(s ) in Mature Protein S53 Glycosite(s) Annotated Protein Sequence >tr|Q5LHV3|Q5LHV3_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF0522 PE=4 SV=1 MKRLIPILLAVFAFAACEKDPDMDKLDNDYLVYTNYDKKADFKQFSTYYIPDS*(53)VLVIGDK KDPEYWKGEAAEAIINAYKENLNSKGFTYTDNKDAADLGIQVSYVQSTYYFTDYGQPEWW WNYPGYWDAPYWGNWGGWYYPYVVNYSITTNSFLTEIMNLKAPEGEKQKLPVLWSSFLSG PASYSGKVNQTLVVRAINQSFAQSPYLTNK Sequence Around Glycosites (21 AA) KQFSTYYIPDSVLVIGDKKDP Glycosite Sequence Logo seqlogo Glycosite Sequence Logo Technique(s) used for Glycosylation Detection Mass shift detected on SDS-polyacrylamide gel and Western blot (using wild-type and mutant B. fragilis cells for expression, and anti-His tag antibody detection) 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