| Project/Area Number |
01480298
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hematology
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| Research Institution | Himeji Institute of Technology (1991)
Niigata University (1989-1990) |
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Principal Investigator |
KOIDE Takehiko Himeji Institute of Tech., Dept. of Life Science, Professor, 理学部, 教授 (60018695)
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| Co-Investigator(Kenkyū-buntansha) |
TOKUNAGA Fuminori Himeji Institute of Tech., Dept. of Life Science, Research Associate, 理学部, 助手 (00212069)
WAKABAYASHI Sadao Himeji Institute of Tech., Dept. of Life Science, Associate Professor, 理学部, 助教授 (80148436)
高橋 芳右 新潟大学, 医学部付属病院, 助手 (70163285)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1989: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Thrombosis / Protein C / Protein C deficiency / Polymorphism / Gene analysis / Histidine-rich glycoprotein / Protein C inhibitor / Chromosome 3 / プロテインC異常症 / HRG / PCR / PCR法 |
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| Research Abstract |
1. Analysis of Genetic Abnormalities for Protein C Deficiency. Makinal use of the polymerase chain reaction (PCR), we analyzed the genes for protein C from Japanese patients with a hereditary protein C deficiency, and identified two molecular abnormalities in two independent probands. One is a 3414T to C mutation in exon VI which converts Tyrl24 (ZAC) to His (CAC) in the second EGF domain. The other is a frameshift deletion of a single G nucleotide in a region of 8854G-8857G in exon IX coding for the carboxy-terminal region, which is predicted to result in an alteration and elongation of the deduced carboxy-terminal amino acid sequence.
2. Diagnosis of Familial Protein C Deficiency. Based on a result described above, we have established a direct diagnostic method of familial protein C deficiency in the second case of a frameshift deletion by use of mutacenic primers for PCR to introduce Ava I or Bal I site into the amplified product.
3. Polymorphism in the Protein C Gene. We have identified two sequence variations in the protein C gene. The first one is an A-T sequence polymorphism at nucleotide -1476 in the untranslated exon 1, and the second one is a T-G sequence polymorphism in the triplet coding for Ser-99 (TCT - TCG).
4. Heparin-Binding Property of Protein C. We found that protein C, and more strongly activated protein C, interacts with heparin, which is enhanced in the presence of calcium ions, and proposed a specific region of protein C as a heparin-binding site.
5. Modulation of Protein C Inhibitor Activity by Histidine-Rich Glycoprotein (HRG). We have found that HRG neutralizes the heparin-dependent inhibitions of activated protein C and thrombin by protein C inhibitor at physioloical concentrations of zinc and calcium ions in plasma, suggesting a new function of HRG as a physiological modulator of protein C inhibitor.
6. Assignment of the Gene for HRG to Chromosome 3. We have assigned the gene for HRG to chromosome 3.
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