ProGlyProt ID BC109 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 BF2494 NCBI Gene ID 3287481 GenBank Gene Sequence 3287481 Protein Information Protein Name BF2494 UniProtKB/SwissProt ID Q5LCH1 NCBI RefSeq YP_212118.1 EMBL-CDS CAH08194.1 UniProtKB Sequence >tr|Q5LCH1|Q5LCH1_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF2494 PE=4 SV=1 MVLLMAVSFAFAQEKNVKEAKSIAGEVKPDFAKAEQLINEALTNPETKDNAATWDVAGYI QKRINEKEMENAYLRKPYDTLKVYNSVLNMYNYYVKCDELAQIPNEKGKIKNKYRSANSK TILAERPNLINGGIQYFNLNKNEDALKYFAAYVDAATLPMMEKENLLEKDTILPQVAYYA TLAADRVGDKDAVMKYAQYALKDKENGQFAMQLLTDAYKAKGDTAKWVEKLQEGIVKFPE NQYFFANLVDYYSSSNQNDKAMQFADDMLAKDPNNKLYLYVKAYLYHNMKDYEKAIEFYK KTLDIDPAYAEACSNLGLVYLLQAQEYADKAPADINDPNYATAQAEIKKFYEAAKPYYEK ARELKPDQKDLWLQGLYRVYYNLNMGPEFEEIEKMM Sequence length 396 AA Subcellular Location Periplasm Function Contains tetratricopeptide repeat (TPR) domains that mediate protein-protein interactions. Protein Structure PDB ID Glycosylation Status Glycosylation Type O (Thr) linked Experimentally Validated Glycosite(s) in Full Length Protein Translation of the protein seems to occur seven codons upstream of the start codon (Met) with a predicted signal peptide (by LipoP) for periplasmic translocation. Therefore, in full length protein sequence the glycosylated residue position should be T87, T178 and T231. Experimentally Validated Glycosite(s ) in Mature Protein T80, T171, T224 Glycosite(s) Annotated Protein Sequence >tr|Q5LCH1|Q5LCH1_BACFN Putative uncharacterized protein OS=Bacteroides fragilis (strain ATCC 25285 / NCTC 9343) GN=BF2494 PE=4 SV=1 MVLLMAVSFAFAQEKNVKEAKSIAGEVKPDFAKAEQLINEALTNPETKDNAATWDVAGYI QKRINEKEMENAYLRKPYDT*(80)LKVYNSVLNMYNYYVKCDELAQIPNEKGKIKNKYRSANSK TILAERPNLINGGIQYFNLNKNEDALKYFAAYVDAATLPMMEKENLLEKDT*(171)ILPQVAYYA TLAADRVGDKDAVMKYAQYALKDKENGQFAMQLLTDAYKAKGDT*(224)AKWVEKLQEGIVKFPE NQYFFANLVDYYSSSNQNDKAMQFADDMLAKDPNNKLYLYVKAYLYHNMKDYEKAIEFYK KTLDIDPAYAEACSNLGLVYLLQAQEYADKAPADINDPNYATAQAEIKKFYEAAKPYYEK ARELKPDQKDLWLQGLYRVYYNLNMGPEFEEIEKMM Sequence Around Glycosites (21 AA) ENAYLRKPYDTLKVYNSVLNM MEKENLLEKDTILPQVAYYAT LTDAYKAKGDTAKWVEKLQEG 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 2009