| Co-Investigator(Kenkyū-buntansha) |
TAKEYA Hiroyuki The Japan Society for the Promotion of Science, JSPS Fellow, 特別研究員(DC) (60222105)
SUEISHI Katsuo Facilty of Mecidine, Professor, Kyushu University, 医学部, 教授 (70108710)
TOKUNAGA Fuminori The Japan Society for the Promotion of Science, JSPS Fellow, 特別研究員(PD) (00212069)
KAWABATA Shun-ichiro Faculty of Science, Research Associate, Kyushu University, 理学部, 助手 (90183037)
MIYATA Toshiyuki Faculty of Science, Research Associate, Kyushu University, 理学部, 助手 (90183970)
ICNINOSE Akitada Department of Biochemistry, Associate Professor, University of Washington, 生化学部, 準教授
DAVIE Earl . W Department of Biochemistry, Professor, University of Washington, 生化学部, 教授
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| Budget Amount *help |
¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 1990: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1989: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1988: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Research Abstract |
The research progress of this year was as follows :
1. We reported the presence of a new trisaccharide composed of two xylose and reducing terminal glucose residues linked to serine residues of bovine blood clotting factors VII and IX (Hase, S., Kawabata, S., Nishimura H., Takeya, H., Sueyoshi, T., Miyata, T., Iwanaga, S., Takao, T., Shimonishi, Y., and Ikenaka, T. (1988) J. Biochem. (Tokyo) 104, 867-868). We describe here the detailed structural analysis of the trisaccharide. Glycopeptides were prepared from bovine factor IX by digestion with Pronase followed by purification by column chromatography. The trisaccharide was released from the protein by the beta-elimination reaction with hydrazine, and the reducing end of the sugar chain was tagged with 2-aminopyridine. The fluorescent pyridylamino derivative of the trisaccharide was purified by gel filtration and reversed-phase high performance liquid chromatography. The glycopeptides and pyridylamino-trisaccharide thus obtained were sub
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jected to methylation study. 500-MHz ^1H nuclear magnetic resonance spectroscopy, and periodate oxidation. Glucose and xylose belong to the D series by high performance liquid chromatography on a chiral column. From the results, the structure of the trisaccharide is proposed as : D-Xylpalpha1-3-D-Xylpalpha-1-3-D-Glcpbeta1-O-Ser-53.
2. Protein Z is a vitamin K-dependent glycoprotein isolated and characterized from human and bovine plasma. A cDNA coding for human protein Z has been obtained by the isolation of phage clones from a liver cDNA library and in vivo amplification of two other liver libraries. Protein Z is synthesized with a prepro-leader sequence of 40 amino acids. The mature protein is composed of 360 residues including a Gla domain of 13 carboxyglutamic acid residues, two epidermal growth factor domains, and a carboxyl terminal region which is highly homologous to the catalytic domain of serine proteases. Human protein Z, however, contains an Asp instead of Ser and a Lys instead of His in the catalytic triad of the active site. Less
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