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


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.

SUBMITTER: Yagi H 

PROVIDER: S-EPMC4030830 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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

Yagi Hirokazu H   Ohno Erina E   Kondo Sachiko S   Yoshida Atsuhiro A   Kato Koichi K  

Biomolecules 20111214 1


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  ...[more]

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