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Glycobiology Advance Access originally published online on June 15, 2005
Glycobiology 2005 15(10):1051-1060; doi:10.1093/glycob/cwi092
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oupjournals.org

Development of structural analysis of sulfated N-glycans by multidimensional high performance liquid chromatography mapping methods

Hirokazu Yagi2,3, Noriko Takahashi2,3,4, Yoshiki Yamaguchi2,3, Naoko Kimura5, Kenji Uchimura6, Reiji Kannagi3,5 and Koichi Kato1,2,3,4

2 Graduate School of Pharmaceutical Sciences, Nagoya City University, 3–1 Tanabe-dori, Mizuho-ku, Nagoya 467–8603, Japan; 3 CREST, JST, 4-1-8 Honcho, Kawaguchi 332–1102, Japan; 4 GLYENCE Co., Ltd. 1-4-6 Masaki, Naka-ku, Nagoya 460–8690, Japan; 5 Program of Molecular Pathology, Aichi Cancer Center, Research Institute, 1–1 Kanokoden, Chikusa-ku, Nagoya 464–8681, Japan; and 6 Department of Anatomy and Program in Immunology, University of California, San Francisco, CA 94143–0452, USA

1 To whom correspondence should be addressed; e-mail: kkato{at}phar.nagoya-cu.ac.jp

Received on May 10, 2005; revised on June 6, 2005; accepted on June 8, 2005

Although the biological importance of sulfated oligosaccharides has been widely recognized, there are only a few reports that describe detailed structures of sulfated N-glycans. This is largely due to the lack of a convenient method to identify structures of sulfated glycans found in low incidence. Here we develop multidimensional high performance liquid chromatography (HPLC) mapping methods for rapid and convenient identification of sulfated N-glycans. By using adequate quantities of sulfated N-glycans derived from LS12 cells, which are transfected with sulfotransferase cDNA, 40 different sulfated glycans have been successfully mapped. Furthermore, we have applied the HPLC data to identification of isomeric products resulting from an enzymatic reaction of N-acetylglucosamine 6-O-sulfotransferase-1 in vitro and revealed that this enzyme preferentially catalyzes sulfation of the GlcNAcß1->2Man{alpha}1->3Man branch in a biantennary acceptor.

Key words: branch specificity / GlcNAc6ST-1 / HPLC mapping / N-glycans / sulfated oligosaccharides


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[Abstract] [Full Text] [PDF]


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