| Project/Area Number |
11470029
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | Hirosaki University |
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Principal Investigator |
ENDO Masahiko Hirosaki University, Department of Biochemistry, Professor, 医学部, 教授 (20004616)
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| Co-Investigator(Kenkyū-buntansha) |
IWAFUNE Mito Hirosaki University, Department of Biochemistry, Research Assistant, 医学部, 助手 (80312487)
KAKIZAKI Ikuo Hirosaki University, Department of Biochemistry, Research Assistant, 医学部, 助手 (80302024)
TAKAGAKI Keiichi Hirosaki University, Department of Biochemistry, Associate Professor, 医学部, 助教授 (70163160)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 2001: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1999: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | proteoglycan / sugar chain reconstruction / glycotechnology / recombinant protein / endo-type glycosidase / endo-β -xylosidase / hyaluronidase / sugar chain transfer reaction / エンド-β-グルクロニダーゼ / グリコサミノグリカン / キシロシル-MU-オリゴ糖 / リコンビナントプロテイン / 糖転移反応 |
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| Research Abstract |
To cover the carbohydrate chain deficiency of reconstructed protein produced by gene engineering, the methods for the reconstruction of glycosaminoglycan (GAG) chains and the introduction of the chains into the recombinant protein using the sugar chain transfer reaction of endo-type glycosidases were investigated, and the following results were obtained.
1. A method, for reconstructing GAG chains according to design was established. The natural GAG chains having no physiological function as donors, and pyridylaminated hexasaccharides prepared from the natural GAG as acceptors, and testicular hyaluronidase were incubated (in Tris-HCl buffer pH 7.0 at 37℃). Systematic combinations of each of the donors and acceptors were used to synthesize reconstructed GAG chains.
2. An oligosaccharide Gal-Gal-xyl-(4-methyiumbelliferone), effective as a carrier to introduce the reconstructed GAG chain into a recombinant protein was discovered. The reconstructed GAG chain bound to the oligosaccharide was introduced into a recombinant protein using synthesized endo-β-xylosidase.
Therefore, a new avenue for glycotechnology was opened.
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