A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows

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...

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Autores principales:	Mehul B. Ganatra, Vladimir Potapov, Saulius Vainauskas, Anthony Z. Francis, Colleen M. McClung, Cristian I. Ruse, Jennifer L. Ong, Christopher H. Taron
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fc15c2b3b3cb4874bff6afec7d990c47
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spelling	oai:doaj.org-article:fc15c2b3b3cb4874bff6afec7d990c472021-12-02T11:46:00ZA bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows10.1038/s41598-020-80488-72045-2322https://doaj.org/article/fc15c2b3b3cb4874bff6afec7d990c472021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80488-7https://doaj.org/toc/2045-2322Abstract 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.Mehul B. GanatraVladimir PotapovSaulius VainauskasAnthony Z. FrancisColleen M. McClungCristian I. RuseJennifer L. OngChristopher H. TaronNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mehul B. Ganatra Vladimir Potapov Saulius Vainauskas Anthony Z. Francis Colleen M. McClung Cristian I. Ruse Jennifer L. Ong Christopher H. Taron A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
description	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.
format	article
author	Mehul B. Ganatra Vladimir Potapov Saulius Vainauskas Anthony Z. Francis Colleen M. McClung Cristian I. Ruse Jennifer L. Ong Christopher H. Taron
author_facet	Mehul B. Ganatra Vladimir Potapov Saulius Vainauskas Anthony Z. Francis Colleen M. McClung Cristian I. Ruse Jennifer L. Ong Christopher H. Taron
author_sort	Mehul B. Ganatra
title	A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
title_short	A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
title_full	A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
title_fullStr	A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
title_full_unstemmed	A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows
title_sort	bi-specific lectin from the mushroom boletopsis grisea and its application in glycoanalytical workflows
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/fc15c2b3b3cb4874bff6afec7d990c47
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A bi-specific lectin from the mushroom Boletopsis grisea and its application in glycoanalytical workflows

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