| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Research Abstract |
Gastrointestinal mucins having GlcNAcα1,4Galβ-R are known to play protective roles against Helicobacter pylori infection. Our research is to develop a synthetic root of artificial mucin with a polyglutamic acid backbone carrying multivalent glycan, which functions as an inhibitor against H pylori infection.
1. Artificial. mucin
Mucins, highly glycosylated proteins that form the major component of mucosa, are known to play a physiological role as barrier to biomolecules. Highly water-soluble, artificial glycopolypeptides with a polyglutamic acid backbone carrying multivalent GlcNAcα1,4Galβ-units, termed 'artificial mucins' have been chemoenzymatically synthesized as potential polymeric inhibitors of infection by H. pylori. In the case, α1,4-N-acetylglucosaminyltransferase (α4GnT) in horse serum, which was obtained by screening of the enzyme activity in various kinds of serums, was used as a tool for synthesizing an epitope of GlcNAc□, 4Gal□-R glycoside.
2. Purification of α4GnT from horse serum
An α4GnT from horse serum was purified 5,300 fold by fractionation with ammonium sulfate, foolowed by sequential chromatographies by CM-Sepharose, Concanavalin A-agarose, Superdex 200 p.g, and UDP-hexanolamine agarose. The purified enzyme was highly specific for an acceptor substrate Galβ-3GalNAcαpNP having core 1 strucrure, on G1cNAc transfer from UDP-G1cNAc donor
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