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Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides

Glycosylation improves the solubility and stability of proteins, contributes to the structural integrity of protein functional sites, and mediates biomolecular recognition events involved in cell-cell communications and viral infections. The first step toward understanding the molecular mechanisms u...

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Detalles Bibliográficos
Autores principales: Yagi, Hirokazu, Ohno, Erina, Kondo, Sachiko, Yoshida, Atsuhiro, Kato, Koichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4030830/
https://www.ncbi.nlm.nih.gov/pubmed/24970123
http://dx.doi.org/10.3390/biom1010048
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author Yagi, Hirokazu
Ohno, Erina
Kondo, Sachiko
Yoshida, Atsuhiro
Kato, Koichi
author_facet Yagi, Hirokazu
Ohno, Erina
Kondo, Sachiko
Yoshida, Atsuhiro
Kato, Koichi
author_sort Yagi, Hirokazu
collection PubMed
description Glycosylation improves the solubility and stability of proteins, contributes to the structural integrity of protein functional sites, and mediates biomolecular recognition events involved in cell-cell communications and viral infections. The first step toward understanding the molecular mechanisms underlying these carbohydrate functionalities is a detailed characterization of glycan structures. Recently developed glycomic approaches have enabled comprehensive analyses of N-glycosylation profiles in a quantitative manner. However, there are only a few reports describing detailed O-glycosylation profiles primarily because of the lack of a widespread standard method to identify O-glycan structures. Here, we developed an HPLC mapping method for detailed identification of O-glycans including neutral, sialylated, and sulfated oligosaccharides. Furthermore, using this method, we were able to quantitatively identify isomeric products from an in vitro reaction catalyzed by N-acetylglucosamine-6O-sulfotransferases and obtain O-glycosylation profiles of serum IgA as a model glycoprotein.
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spelling pubmed-40308302014-06-24 Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides Yagi, Hirokazu Ohno, Erina Kondo, Sachiko Yoshida, Atsuhiro Kato, Koichi Biomolecules Article Glycosylation improves the solubility and stability of proteins, contributes to the structural integrity of protein functional sites, and mediates biomolecular recognition events involved in cell-cell communications and viral infections. The first step toward understanding the molecular mechanisms underlying these carbohydrate functionalities is a detailed characterization of glycan structures. Recently developed glycomic approaches have enabled comprehensive analyses of N-glycosylation profiles in a quantitative manner. However, there are only a few reports describing detailed O-glycosylation profiles primarily because of the lack of a widespread standard method to identify O-glycan structures. Here, we developed an HPLC mapping method for detailed identification of O-glycans including neutral, sialylated, and sulfated oligosaccharides. Furthermore, using this method, we were able to quantitatively identify isomeric products from an in vitro reaction catalyzed by N-acetylglucosamine-6O-sulfotransferases and obtain O-glycosylation profiles of serum IgA as a model glycoprotein. MDPI 2011-12-14 /pmc/articles/PMC4030830/ /pubmed/24970123 http://dx.doi.org/10.3390/biom1010048 Text en © 2011 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Yagi, Hirokazu
Ohno, Erina
Kondo, Sachiko
Yoshida, Atsuhiro
Kato, Koichi
Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title_full Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title_fullStr Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title_full_unstemmed Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title_short Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
title_sort development and application of multidimensional hplc mapping method for o-linked oligosaccharides
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4030830/
https://www.ncbi.nlm.nih.gov/pubmed/24970123
http://dx.doi.org/10.3390/biom1010048
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