Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex

Abstract Lectins are proteins with the ability to recognize and bind to specific glycan structures. These molecules play important roles in many biological systems and are actively being studied because of their ability to detect glycan biomarkers for many diseases. Hemagglutinin (HA) proteins from...

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Autores principales:	Ea Kristine Clarisse Tulin, Chiaki Nakazawa, Tomomi Nakamura, Shion Saito, Naoki Ohzono, Keiko Hiemori, Shin-ichi Nakakita, Hiroaki Tateno, Takashi Tonozuka, Atsushi Nishikawa
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cc0966aae15445da915caa89bf072130
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spelling	oai:doaj.org-article:cc0966aae15445da915caa89bf0721302021-11-14T12:21:06ZGlycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex10.1038/s41598-021-01501-12045-2322https://doaj.org/article/cc0966aae15445da915caa89bf0721302021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01501-1https://doaj.org/toc/2045-2322Abstract Lectins are proteins with the ability to recognize and bind to specific glycan structures. These molecules play important roles in many biological systems and are actively being studied because of their ability to detect glycan biomarkers for many diseases. Hemagglutinin (HA) proteins from Clostridium botulinum type C neurotoxin complex; HA1, HA2, and HA3 are lectins that aid in the internalization of the toxin complex by binding to glycoproteins on the cell surface. HA1 mutants have been previously reported, namely HA1 W176A/D271F and HA1 N278A/Q279A which are specific to galactose (Gal)/N-acetylgalactosamine (GalNAc) and N-acetylneuraminic acid (Neu5Ac) sugars, respectively. In this study, we utilized HA1 mutants and expressed them in complex with HA2 WT and HA3 WT to produce glycan detecting tools with high binding affinity. Particularly, two types were made: Gg and Rn. Gg is an Alexa 488 conjugated lectin complex specific to Gal and GalNAc, while Rn is an Alexa 594 conjugated lectin complex specific to Neu5Ac. The specificities of these lectins were identified using a glycan microarray followed by competitive sugar inhibition experiments on cells. In addition, we confirmed that Gg and Rn staining is clearly different depending on cell type, and the staining pattern of these lectins reflects the glycans present on the cell surface as shown in enzyme treatment experiments. The availability of Gg and Rn provide us with new promising tools to study Gal, GalNAc, and Neu5Ac terminal epitopes which can aid in understanding the functional role of glycans in physiological and pathological events.Ea Kristine Clarisse TulinChiaki NakazawaTomomi NakamuraShion SaitoNaoki OhzonoKeiko HiemoriShin-ichi NakakitaHiroaki TatenoTakashi TonozukaAtsushi NishikawaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ea Kristine Clarisse Tulin Chiaki Nakazawa Tomomi Nakamura Shion Saito Naoki Ohzono Keiko Hiemori Shin-ichi Nakakita Hiroaki Tateno Takashi Tonozuka Atsushi Nishikawa Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
description	Abstract Lectins are proteins with the ability to recognize and bind to specific glycan structures. These molecules play important roles in many biological systems and are actively being studied because of their ability to detect glycan biomarkers for many diseases. Hemagglutinin (HA) proteins from Clostridium botulinum type C neurotoxin complex; HA1, HA2, and HA3 are lectins that aid in the internalization of the toxin complex by binding to glycoproteins on the cell surface. HA1 mutants have been previously reported, namely HA1 W176A/D271F and HA1 N278A/Q279A which are specific to galactose (Gal)/N-acetylgalactosamine (GalNAc) and N-acetylneuraminic acid (Neu5Ac) sugars, respectively. In this study, we utilized HA1 mutants and expressed them in complex with HA2 WT and HA3 WT to produce glycan detecting tools with high binding affinity. Particularly, two types were made: Gg and Rn. Gg is an Alexa 488 conjugated lectin complex specific to Gal and GalNAc, while Rn is an Alexa 594 conjugated lectin complex specific to Neu5Ac. The specificities of these lectins were identified using a glycan microarray followed by competitive sugar inhibition experiments on cells. In addition, we confirmed that Gg and Rn staining is clearly different depending on cell type, and the staining pattern of these lectins reflects the glycans present on the cell surface as shown in enzyme treatment experiments. The availability of Gg and Rn provide us with new promising tools to study Gal, GalNAc, and Neu5Ac terminal epitopes which can aid in understanding the functional role of glycans in physiological and pathological events.
format	article
author	Ea Kristine Clarisse Tulin Chiaki Nakazawa Tomomi Nakamura Shion Saito Naoki Ohzono Keiko Hiemori Shin-ichi Nakakita Hiroaki Tateno Takashi Tonozuka Atsushi Nishikawa
author_facet	Ea Kristine Clarisse Tulin Chiaki Nakazawa Tomomi Nakamura Shion Saito Naoki Ohzono Keiko Hiemori Shin-ichi Nakakita Hiroaki Tateno Takashi Tonozuka Atsushi Nishikawa
author_sort	Ea Kristine Clarisse Tulin
title	Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
title_short	Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
title_full	Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
title_fullStr	Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
title_full_unstemmed	Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex
title_sort	glycan detecting tools developed from the clostridium botulinum whole hemagglutinin complex
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/cc0966aae15445da915caa89bf072130
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Glycan detecting tools developed from the Clostridium botulinum whole hemagglutinin complex

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