1997 Fiscal Year Final Research Report Summary
A novel serine/threonine kinase associated with thrombin receptor signaling in human platelet
| Project/Area Number |
08680680
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Mie University |
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Principal Investigator |
IDO Masaru Mie University Faculty of Medicine, Lecturer, 医学部, 講師 (90167263)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Koji Mie University Faculty of Medicine, Professor, 医学部, 教授 (70077808)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Thrombin receptor / Protein kinase / Platelet |
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| Research Abstract |
Novel protein kinases that may associate with thrombin receptor were sought by a procedure based on the ability of these enzymes to undergo renaturation and catalyze the phosphorylation of a protein substrate fixed in a gel. We employed a fusion protein comprising of glutathione S-transferase and the substrate protein that corresponds to residues 375-421 of thrombin receptor (GST-TRCD). We report that platelets contain a serine/threonine protein kinase with molecular masse of about 33 kDa, which specifically bound to GST-TRCD.Treatment of platelets with thrombin increased the thrombin receptor-associated kinase (TRAK) activity by 700%. TRAK was also activated by a thrombin receptor agonist peptide, but not by hirudin-treated or diisopropylphosphate-inactivated thrombin. Activation of thrombin receptor resulted in phosphorylation of TRCD by TRAK in a Ca^<2+>-dependent way. In summary, our results provide evidence of a novel ligand-activated serine/threonine kinase that associates with the cytoplasmic domain of the thrombin receptor.
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[Publications] Deguchi, H., Takeya, H., Gabazza, E.C., Hayashi, N., and Suzuki, K.: "Dilazep, an adenosine potentiator, inhibits tissue factor expression in the endothelial cells stimulated with tumor necrosis factor-alpha, thrombin or phorbo1 12-myristate 13-acetate." Blood. 90. 2345-2356 (1997)
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[Publications] Takeya, H., Mori, T., Gabazza, E.C., Kuroda, K., Deguchi, H., Matsuura, E., Ichikawa, K., Koike, T.and Suzuki, K.: "Anti-beta2-glycoprotein I (beta2GPI) monoclonal antibodies with lupus anticoagulant-like activity enhance the beta2GPI binding to phospholipids." J.Clin.Invest.99. 2260-2268 (1997)
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[Publications] Gabazza, E.C., Takeya, H., Deguchi, H., Taguchi, O., Wada, H., Nishioka, J., Zhou, H.and Suzuki, K.: "Aquired activated protein C resistance in patients with lupus anticoagulant and essential thrombocythemia." Clin.Appl.Thromb.Hemostas.3. 119-123 (1997)
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[Publications] Ido, M., Ogawa, H., Kato, S., Hayashi, K., and Suzuki, K.: A novel serine/threonine kinase associated with thrombin receptor signaling in human platelets. In : Blood Coagulation, Fibrinolysis, and Platelets. Davie EW,Kakeshita K,eds.Splinger-Verlag Tokyo, 113-6 (1996)
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