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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2011
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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. |
format | Online Article Text |
id | pubmed-4030830 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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