Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS

Xingyu Zhu; Kenichi Hatanaka; Tatsuya Yamagata; Toshinori Sato

doi:10.1002/elps.200800719

Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS

Xingyu Zhu, Kenichi Hatanaka, Tatsuya Yamagata, Toshinori Sato

Department of Biosciences and Informatics

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

Abstract

A glycosphingolipid analogue (12-azidododecyl β-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl b-lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CEESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.

Original language	English
Pages (from-to)	3519-3526
Number of pages	8
Journal	ELECTROPHORESIS
Volume	30
Issue number	20
DOIs	https://doi.org/10.1002/elps.200800719
Publication status	Published - 2009

Keywords

CE-ESI-MS
Neutral carbohydrate
Saccharide primer

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Clinical Biochemistry

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10.1002/elps.200800719

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title = "Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS",

abstract = "A glycosphingolipid analogue (12-azidododecyl β-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl b-lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CEESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.",

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AU - Zhu, Xingyu

AU - Hatanaka, Kenichi

AU - Yamagata, Tatsuya

AU - Sato, Toshinori

PY - 2009

Y1 - 2009

N2 - A glycosphingolipid analogue (12-azidododecyl β-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl b-lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CEESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.

AB - A glycosphingolipid analogue (12-azidododecyl β-lactoside) as a saccharide primer has been shown to be useful for the synthesis of oligosaccharide libraries by mammalian cells. In the present study, CE-ESI-MS was employed to elucidate the structure of glycosphingolipid analogues derived from 12-azidododecyl b-lactoside (Lac-C12N3) by mammalian cells. MDCK cells and COLO201 cells were cultured with Lac-C12N3, and the glycosylated products secreted into the medium were collected and separated into acidic and neutral products by column chromatography. The acidic products could be directly analyzed by CEESI-MS, while the neutral products were converted to anionic derivatives via a reaction with propiolic acid. With this method, it was possible to analyze both acidic and neutral products glycosylated by MDCK cells and COLO201 cells at high sensitivity.

KW - CE-ESI-MS

KW - Neutral carbohydrate

KW - Saccharide primer

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Structural analysis of glycosphingolipid analogues obtained by the saccharide primer method using CE-ESI-MS

Abstract

Keywords

ASJC Scopus subject areas

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