| Project/Area Number |
01570109
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Saga Medical School |
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Principal Investigator |
ASAKAWA Takeo Saga Medical School, Dept. of Pharmacol. Prof., 医学部, 教授 (50028362)
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| Co-Investigator(Kenkyū-buntansha) |
TAKANO Masako Saga Medical School, Dept. of Pharmacol. Assist. Prof., 医学部, 助手 (00154807)
ENOMOTO Keiichi Saga Medical School, Dept. of Pharmacol. Assist. Prof., 医学部, 助手 (20128127)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Guanylate cyclase / Unsaturated fatty acid / カルシウム / GTP / グアニレ-トシクラ-ゼ / サイクリックGMP / シナプス膜 |
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| Research Abstract |
1. We found a membrane bound type of guanylate cyclase activated by Ca^<2+> and unsaturated fatty acid in synaptic plasma membranes. The activation of the cyclase required co-existence of GTP or its analogues. The cyclase was tried to be purified, under retaining the properties of being activated by Ca^<2+>, GTP and unsaturated fatty acid. After several steps of chromatographies ; ion exchange, GTP-affinity, hydrophobic affinity, WGA-affinity and gel filtration, the highly purified cyclase was obtained. The finally purified enzyme was observed to be stimulated by the activators above, but in less extent than that in the membranes.
2. The binding of Ca^<2+>, GTP and unsaturated fatty acid to bands in acrylamide gels from SDS-PAGE with the finally purified enzyme was examined to identify sub-units of the enzyme which participated in the activation.
3. Kinetic properties of the activation by GTP, Ca^<2+> and unsaturated fatty acid were analyzed by using computer programs.
4. Among several chemical compounds tested, a group of compounds was found to inhibit the cyclase selectively. This finding is useful in elucidating a physiological role of this cyclase in cell function.
5. Specificity of unsaturated fatty acid in the activation was studied. Unsaturated fatty acid-peroxide was observed to be ineffective.
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