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A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
- Source :
- Scientific Reports, Scientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
- Publication Year :
- 2021
- Publisher :
- Springer Science and Business Media LLC, 2021.
-
Abstract
- The BLL lectin from the edible Japanese “Kurokawa” mushroom (Boletopsis leucomelaena) was previously reported to bind to N-glycans harboring terminal N-acetylglucosamine (GlcNAc) and to induce apoptosis in a leukemia cell line. However, its gene has not been reported. In this study, we used a transcriptomics-based workflow to identify a full-length transcript of a BLL functional ortholog (termed BGL) from Boletopsis grisea, a close North American relative of B. leucomelaena. The deduced amino acid sequence of BGL was an obvious member of fungal fruit body lectin family (Pfam PF07367), a highly conserved group of mushroom lectins with a preference for binding O-glycans harboring the Thomsen–Friedenreich antigen (TF-antigen; Galβ1,3GalNAc-α-) and having two ligand binding sites. Functional characterization of recombinant BGL using glycan microarray analysis and surface plasmon resonance confirmed its ability to bind both the TF-antigen and β-GlcNAc-terminated N-glycans. Structure-guided mutagenesis of BGL’s two ligand binding clefts showed that one site is responsible for binding TF-antigen structures associated with O-glycans, whereas the second site specifically recognizes N-glycans with terminal β-GlcNAc. Additionally, the two sites show no evidence of allosteric communication. Finally, mutant BGL proteins having single functional bindings site were used to enrich GlcNAc-capped N-glycans or mucin type O-glycopeptides from complex samples in glycomics and glycoproteomics analytical workflows.
- Subjects :
- Proteomics
0301 basic medicine
Glycan
Glycosylation
Science
Mutant
Article
Fungal Proteins
Glycomics
Fungal biology
03 medical and health sciences
Polysaccharides
Lectins
Humans
Amino Acid Sequence
Peptide sequence
Boletopsis grisea
Binding Sites
Multidisciplinary
030102 biochemistry & molecular biology
biology
Chemistry
Basidiomycota
Lectin
carbohydrates (lipids)
030104 developmental biology
Biochemistry
Fungal fruit body lectin family
biology.protein
Medicine
Boletopsis leucomelaena
Agaricales
Sequence Alignment
Protein Binding
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....9eeaffc0523e710a13b23ee2173a9a27