Roles of G-proteins and cytoskeletal porteins in transmitter release.

G 蛋白和细胞骨架蛋白在递质释放中的作用。

基本信息

批准号：
05680683
负责人：
KUMAKURA Konosuke
金额：
$ 1.28万
依托单位：
Sophia University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680683/
关键词：
Mechanism of Exocytosis Transmitter Release GTP-Binding Proteins Myosin Light Chain Kinase Ca^<2+>-Dependent Release ATP-Dependent Priming Mastoparan Cytoskeletal Proteins ヴォルトマニン

项目摘要

The aim of this research project is to clarify the roles of GTP-binding proteins and cytoskeletal proteins in the mechanism of transmitter release.Previously we have suggested that Go, one of heterotrimeric GTP-binding proteins, negatively regulates Ca^<2+>-dependent release of catecholamine in permeabilized chromaffin cells.We also have suggested a possible involvement of myosin light chain kinase in the mechanism of Ca^<2+>-dependent exocytosis.In the project, we attempted to elucidate the mechanism whereby Ca^<2+>-dependent release is regulated by Go, and by myosin light chain kinase in permeabilized chromaffin cells.We have obtained the following results.1.By use of mastoparan, a peptide known to activate directly GTP-binding proteins, and by use of anti-Go antibodies, we found that the negative regulation by Go is on the process of ATP-dependent priming of exocytosis.This inhibitory Go was suggested to be activated by GAP43, and activation of protein kinase C seemed to stimulate Ca^<2+>-dependent release by abolishing the action of GAP43 on Go.2.We found that myosin light chain kinase is essential for ATP-dependent priming of exocytosis, by use of specific inhibitors of myosin light chain kinase, wortmannin and SM-1.Histochemical studies suggested that the inhibition of myosin light chain kinase inhibits reorganization of cortical F-actin, and thus suggested an important role of cortical F-actin for priming of chromaffin vesicles to the exocytotic site.

The aim of this research project is to clarify the roles of GTP-binding proteins and cytoskeletal proteins in the mechanism of transmitter release.Previously we have suggested that Go, one of heterotrimeric GTP-binding proteins, negatively regulates Ca^<2+>-dependent release of catecholamine in permeabilized chromaffin cells.We also have suggested a possible involvement of肌球蛋白光链激酶在Ca^<2+> - 依赖性胞吐作用的机制中。在项目中，我们试图阐明Ca^<2+> - 依赖性释放受GO调节的机制，并由肌球蛋白轻链激酶在渗透的成年蛋白中的肌球蛋白轻链激酶中得到了以下结果。蛋白质以及使用抗Go抗体，我们发现，GO的负调节是在依赖ATP的依赖ATP的外吞作用的过程中。建议这种抑制性GO被GAP43激活，并且激活蛋白激酶C的激活，蛋白激酶C的激活似乎刺激了Ca^<2+> - 依赖于iSINDEL ISIS ISSIS of gop43 on goap43 on goap43 on gop43。 ATP-dependent priming of exocytosis, by use of specific inhibitors of myosin light chain kinase, wortmannin and SM-1.Histochemical studies suggested that the inhibition of myosin light chain kinase inhibits reorganization of cortical F-actin, and thus suggested an important role of cortical F-actin for priming of chromaffin vesicles to the exocytotic site.