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Glycobiology Advance Access published online on June 15, 2005
Glycobiology, 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
Received May 10, 2005
Revised June 6, 2005
Accepted June 8, 2005

Article

Development of structural analysis of sulfated N-glycans by multi-dimensional HPLC mapping methods

Hirokazu Yagi 1, Noriko Takahashi 2, Yoshiki Yamaguchi 1, Naoko Kimura 3, Kenji Uchimura 4, Reiji Kannagi 5, and Koichi Kato 2*

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

* To whom correspondence should be addressed.
Koichi Kato, E-mail: kkato{at}phar.nagoya-cu.ac.jp


  Abstract

Although 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 the sulfated glycans with a low incidence. Here we develop multi-dimensional HPLC mapping methods for a 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 GlcNAc{beta}1->2Man{alpha}1->3Man branch in a bi-antennary acceptor.

Keywords: sulfated oligosaccharides/N-glycans/HPLC mapping/GlcNAc6ST-1/branch specificity.
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