| 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 | Putative fucosyl transferase |
| Predicted Accessory Gene(s) | BF4298-4306 region lfg (locus of fragilis glycosylation). |
| Additional Comment | 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. Ile, Leu, and Val were found most frequently whereas Met is rarest at third position (reflecting the otherwise low number of Mets in proteins compared with the other five amino acids at the third position of the motif). The methyl group-containing amino acid at the third position being unreactive may play a role only in recognition of the site, whereas Asp residue may play a catalytic role. |
| Literature |
| Year of Identification | 2009 |
| Year of Identification Month Wise | 2009.4.17 |
| Year of Validation | 2011 |
| Reference | 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. Journal of Biological Chemistry, 286(5), pp.3219-3226. |
| Corresponding Author | Laurie E. Comstock |
| Contact | Channing Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA. |
| Reference | 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(2), pp.321-331. |
| Corresponding Author | Laurie E. Comstock |
| Contact | Channing Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA. |
| Reference | 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] |
| Author | Fletcher, C.M., Coyne, M.J. and Comstock, L.E. |
| Research Group | Channing Laboratory, Brigham and Womens Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA. |
| Corresponding Author | Comstock, L.E. |
| Contact | Channing Laboratory, Brigham and Womens Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA. |
| Reference | 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] |
| Author | Fletcher, C.M., Coyne, M.J., Villa, O.F., Chatzidaki-Livanis, M. and Comstock, L.E. |
| Research Group | Channing Laboratory, Brigham & Womens Hospital, Harvard Medical School, 181 Longwood Avenue, Boston, MA 02115, USA. |
| Corresponding Author | Comstock, L.E. |
| Contact | Channing Laboratory, Brigham & Womens Hospital, Harvard Medical School, 181 Longwood Avenue, Boston, MA 02115, USA. |
