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Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides
Graduate School of Pharmaceutical Science, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan
Graduate School of Medical Sciences and Medical School, Nagoya City University, Kawasumi-1, Mizuho-cho Mizuho-ku, Nagoya 467-8601, Japan
Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, 5-1 Higashiyama Myodaiji, Okazaki 444-8787, Japan
GLYENCE Co., Ltd., 2-22-8 Chikusa, Chikusa-ku, Nagoya 464-0858, Japan
The Glycoscience Institute, Ochanomizu University, 2-1-1 Ohtsuka, Bunkyo-ku, Tokyo 112-8610, Japan
* Author to whom correspondence should be addressed.
Received: 29 August 2011; in revised form: 30 November 2011 / Accepted: 5 December 2011 / Published: 14 December 2011
Abstract: 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.
Keywords: O-glycans; HPLC map; glycosylation profiling; sulfated oligosaccharide; IgA
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Yagi, H.; Ohno, E.; Kondo, S.; Yoshida, A.; Kato, K. Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides. Biomolecules 2011, 1, 48-62.
Yagi H, Ohno E, Kondo S, Yoshida A, Kato K. Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides. Biomolecules. 2011; 1(1):48-62.
Yagi, Hirokazu; Ohno, Erina; Kondo, Sachiko; Yoshida, Atsuhiro; Kato, Koichi. 2011. "Development and Application of Multidimensional HPLC Mapping Method for O-linked Oligosaccharides." Biomolecules 1, no. 1: 48-62.