Roles of the protein kinases on platelet functions

蛋白激酶对血小板功能的作用

• 批准号: 04671511
• 负责人: HASHIMOTO Yoshiaki
• 金额: $ 1.41万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671511/
• 关键词: platelet aggregation; ATP release; myosin light chain kinase; protein kinase C; wortmannin

We examined the roles of the protein kinases in platelet aggregation and release.<Methods>1.Platelets : Platelet rich plasma and washed platelets were prepared from bloods of healthy volunteers.2.Measurement of platelet responses : Platelet aggregation and ATP release were monitored using a CAF-100 Ca^<2+> analyzer. ATP release was measured by the luciferin-luciferase reaction.<Results>1.The roles of myosin light chain kinase.The specific inhibitor of myosin light chain kinase wortmannin inhibited ADP-induced platelet aggregation with no effect on their shape change.2.The roles of protein kinase C.The specific inhibitor of protein kinase C PKC-I did not affect much U46619-induced platelet aggregation and ATP release. However the inhibition of platelet aggregation with GRGDS prevented ATP release in the presence of PKC-I.On the other hand, ATP release was not seen in the presence of PKC-I in washed platelets.<Conclusion>1.It was suggested that myosin light chain kinase activation is a prerequisite for ADP-induced platelet aggregation, but not for changes in their shape.2.It was suggested that there are protein-kinase C-dependent and -independent mechanisms for ATP release by human platelets and that activation of the latter mechanism may depend on aggregation and plasma factors.

我们检查了蛋白激酶在血小板聚集和释放中的作用。<方法>1.血小板：从健康志愿者的血液中制备富含血小板的血浆和洗涤的血小板。2.血小板反应的测量：使用CAF-100 Ca^2+分析仪监测血小板聚集和ATP释放。通过荧光素-荧光素酶反应测定ATP释放。<结果>1.肌球蛋白轻链激酶的作用。肌球蛋白轻链激酶特异性抑制剂渥曼青霉素抑制ADP诱导的血小板聚集，但对其形状变化没有影响。2.蛋白激酶C的作用。蛋白激酶C特异性抑制剂PKC-I对U46619诱导的血小板聚集影响不大。 血小板聚集和 ATP 释放。然而，GRGDS 抑制血小板聚集，在 PKC-I 存在的情况下阻止了 ATP 的释放。另一方面，在洗涤的血小板中，在 PKC-I 存在的情况下，没有观察到 ATP 的释放。<结论>1. 提示肌球蛋白轻链激酶激活是 ADP 诱导血小板聚集的先决条件，但不是其形状改变的先决条件。 2. 提示存在蛋白激酶 人血小板释放 ATP 的 C 依赖性和非依赖性机制，后一种机制的激活可能取决于聚集和血浆因素。

项目成果

橋本佳明: "受容体作動性チャネルの活性化機構" BIO medica. 8. 416-419 (1993)

Yoshiaki Hashimoto：“受体激动剂通道的激活机制”BIO medica. 8. 416-419 (1993)。

Yoshiaki Hashimoto: "Two thrombin-activated Ca^<2+> channels in human platelets." J.Biol.Chem.267. 17078-17081 (1992)

Yoshiaki Hashimoto：“人类血小板中有两个凝血酶激活的 Ca^2 通道。”

Yoshiaki Hashimoto et al: "Two thrombin-activated Ca^<2+> channels in human platelets" J.Biol Chem. 267. 17078-17081 (1992)

Yoshiaki Hashimoto 等人：“人血小板中的两个凝血酶激活的 Ca^2 通道”J.Biol Chem。

HASHIMOTO,OGIHARA,NAKANISHI,MATSUDA,KUROKAWA,NONOMURA: "Two Thrombin-Activated Ca^<2+> Channels in Human Platelets" J.Biol.Chem.267. 17078-17081 (1992)

HASHIMOTO、OGIHARA、NAKANISHI、MATSUDA、KUROKAWA、NONOMURA：“人血小板中的两个凝血酶激活的 Ca^<2> 通道”J.Biol.Chem.267。

Yoshiaki Hashimoto et al: "Ca^<2+> entry pathways activated by the tumor promoter thapsigargin in human platelets" Biochim.Biophys Acta.1220. 37-41 (1993)

Yoshiaki Hashimoto等人：“人血小板中肿瘤启动子毒胡萝卜素激活的Ca 2+ 进入途径”Biochim.Biophys Acta.1220。