Glycobiology Advance Access published online on October 30, 2002
Glycobiology, doi:10.1093/glycob/cwg001
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© 2002 Oxford University Press
ORIGINAL ARTICLES
1 Department of Biochemistry, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550 Midkine (MK), a heparin-binding growth factor, binds strongly to oversulfated structures in chondroitin sulfates and heparan sulfate. To elucidate the carbohydrate structure actually involved in the strong binding, dissected brains from 13-day mouse embryos were incubated with [14C]-glucosamine. The labeled glycosaminoglycans were fractionated by MK-agarose affinity chromatography to a weakly binding fraction, which was eluted by 0.5 M NaCl, and a strongly binding fraction, which was eluted by higher NaCl concentrations. Among the unsaturated disaccharides released from the strongly binding fraction by chondroitinase ABC,
Revised on August 16, 2002
Accepted on August 18, 2002
Glycosaminoglycan structures required for strong binding to midkine, a heparin binding growth factor
2 Department of Life Science, Aichi University of Education, Kariya, Aichi 448-8542
3 Meiji Dairies Corporation, 540 Naruda, Odawara 250-0862, Japan
Di-diSE with 4,6-disulfated N-acetylgalactosamine comprised 32.3 %, while its content was less in the weakly binding fraction. Artificial chondroitin sulfate E structure was formed using N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase purified from squid or the recombinant human enzyme. Analysis of the products and their interaction with MK revealed that E unit without 3-O-sulfation of glucuronic acid is sufficient for the strong binding, provided that they are present as a dense cluster. Among the sulfated disaccharides released by heparitinase digestion, the tri-sulfated one, DiHS-triS, was the most abundant in the strongly binding fraction, and was less in the weakly binding fraction. Together with results of previous studies, we concluded that the multivalent tri-sulfated heparin-like unit is another structure involved in strong binding to MK.
Keywords: chondroitin sulfate / heparan sulfate / sulfotransferase
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