Binds to N-glycans having at least one terminal outer arm GlcNAc as well as O-glycans containing Gal-β1,3-GalNAc-α- within their structure.
Enables enrichment of GlcNAc-capped N-glycans or mucin type O-glycopeptides from complex samples in glycomics and glycoproteomics analytical workflows.
This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.
Boletopsis grisea Lectin (BGL) is a recombinant 15 kDa lectin from the Boletopsis grisea mushroom that has been expressed in E. coli. BGL has two separately functioning ligand binding sites (1). Site 1 binds to O-glycans bearing the Tn antigen (GalNAc-α-Ser/Thr) or Thomsen-Friedenreich antigen (TF-antigen; Gal-β1,3-GalNAc-α-) and Site 2 binds N-glycans with terminal GlcNAc residues.
Boletopsis grisea Lectin (BGL) binding sites
BGL has two separately functioning ligand binding sites (1). Site 1 binds to O-glycans bearing the Tn antigen (GalNAc-α-Ser/Thr) or Thomsen-Friedenreich antigen (TF-antigen; Gal-β1,3-GalNAc-α-) and Site 2 binds N-glycans with terminal GlcNAc residues.
This product is related to the following categories:
100 µg BGL enriches >85% Procainamide-G0/A2 N-glycan in an Epitope Directed Glycan Enrichment (EDGE) assay (1,2). The assay involves an enrichment protocol termed EDGE whereby 100 µg BGL lectin is incubated with 100 ng Procainamide-G0 N-glycan substrate for 1.5 hours at 25°C in 20 mM Tris-HCl pH 7.5 buffer in a total volume of 120 µl.
BGL has only been tested for the enrichment of O-glycopeptides and free N-glycans in EDGE assays.
Optimal incubation time (60 to 90 minutes) will depend on the specific substrate.
A lower starting amount of BGL for the EDGE assay could be used for O-glycopeptides. This amount should be empirically determined.
The formulation of BGL contains 50 mM Tris-HCl, pH 7.5 and 200 mM NaCl. If this salt concentration will affect downstream workflow (e.g., capillary electrophoresis analysis), buffer exchange treatment of BGL may be performed prior to proceeding with the enrichment reaction.
References
Ganatra, M. B. et al. (2021). A bi‑specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows. Sci. Rep. 11, 160.
Vainauskas, S. et al. (2016). Profiling of core fucosylated N-glycans using a novel bacterial lectin that specifically recognizes α1,6 fucosylated chitobiose. Sci. Rep. 6, 34195.
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