Glycobiology Advance Access published online on February 22, 2007
Glycobiology, doi:10.1093/glycob/cwm021
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Structural determination of novel sulfated octasaccharides isolated from chondroitin sulfate of shark cartilage and their application for characterizing monoclonal antibody epitopes
2 Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan
3 Laboratory of Proteoglycan Signaling and Therapeutics, Graduate School of Life Science, Hokkaido University, Frontier Research Center for Post-Genomic Science and Technology, Nishi 11-choume, Kita 21-jo, Kita-ku, Sapporo, Hokkaido 001-0021, Japan
4 Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Kyoto 603-8558, Japan.
1 To whom corresponding should be addressed at the present address: Laboratory of Proteoglycan Signaling and Therapeutics, Graduate School of Life Science, Hokkaido University, Frontier Research Center for Post-Genomic Science and Technology, Nishi 11-choume, Kita 21-jo, Kita-ku, Sapporo, Hokkaido 001-0021, Japan. Tel.: 81-(11)-706-9054; Fax: 81-(11)-706-9056; E-mail: k-sugar{at}sci.hokudai.ac.jp
Received on October 26, 2006; revised on January 24, 2007; accepted on February 19, 2007
Twelve octasaccharide fractions were obtained from chondroitin sulfate C derived from shark cartilage after hyaluronidase digestion. Their sugar and sulfate composition was assigned by MALDI-TOF/MS. The sequences were determined at low picomole amounts by a combination of enzymatic digestions with HPLC, and were composed of disaccharide building units including O [GlcUAb1-3GalNAc], C [GlcUAb1-3GalNAc(6S)], A [GlcUAb1-3GalNAc(4S)] and/or D [GlcUA(2S)b1-3GalNAc(6S)], where 2S, 4S and 6S represent 2-O-, 4-O- and 6-O-sulfate, respectively. As many as 24 different sequences including minor ones were revealed, exhibiting a high degree of structural diversity reflecting the enormous heterogeneity of the parent polysaccharides. Nineteen of them were novel, with the other four reported previously as unsaturated counterparts obtained after digestion with chondroitinase. A microarray of these structurally defined octasaccharide fractions were prepared using low picomole amounts of their lipid-derivatives to investigate the binding specificity of four commercial anti-chondroitin sulfate antibodies CS-56, MO-225, 2H6 and LY111. The results revealed that multiple unique sequences were recognized by each antibody, which implies that the common conformation shared by the multiple primary sequences in the intact CS chains is important as an epitope for each mAb. Comparison of the specificity of the tested antibodies indicates that CS-56 and MO-225 specifically recognize octasaccharides containing an A-D tetrasaccharide sequence, whereas 2H6 and LY111 require a hexasaccharide as a minimum size for their binding, and prefer sequences with A- and C-units such as C-C-A-C (2H6) or C-C-A-O, C-C-A-A and C-C-A-C (LY111) for strong binding but require no D-unit.
Key words: chondroitin sulfate / sugar sequencing / sulfation / antibody epitope / octasaccharides
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  P. Pothacharoen, K. Kalayanamitra, S. S. Deepa, S. Fukui, T. Hattori, N. Fukushima, T. Hardingham, P. Kongtawelert, and K. Sugahara Two Related but Distinct Chondroitin Sulfate Mimetope Octasaccharide Sequences Recognized by Monoclonal Antibody WF6 J. Biol. Chem., November 30, 2007; 282(48): 35232 - 35246. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Yamada, H. Morimoto, T. Fujisawa, and K. Sugahara Glycosaminoglycans in Hydra magnipapillata (Hydrozoa, Cnidaria): demonstration of chondroitin in the developing nematocyst, the sting organelle, and structural characterization of glycosaminoglycans Glycobiology, August 1, 2007; 17(8): 886 - 894. [Abstract] [Full Text] [PDF]
|
|