Modification mechanisms of single ionic channel currents from cardiac sarcoplasmic reticulum and mitochondria by physiologically active substances.

生理活性物质对心脏肌浆网和线粒体单离子通道电流的修饰机制。

• 批准号: 05680729
• 负责人: UEHARA Akira
• 金额: $ 1.28万
• 依托单位: Fukuoka 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-05680729/
• 关键词: ionic channels; cardiac muscle; sarcoplasmic reticulum; single channel currents; 単一イオンチャネル電流

Effects of physiologically active substances on the single channel molecules from cardiac sarcoplasmic reticulum and mitochondria were examined by using the planar lipid bilayr, the hot ligand binding, and the ^<45>Ca^<2+> efflux methods. Systematic screening works were performed in great many kinds of substances and ionic channels. It is the most important findings that cytoplasmic application of arachidonic acids and polyamines causes a potent functional modulation in the ryanodine receptor channels. We have already published a paper related to the activated mechanism by the arachidonic acid and analogs, the summary of which is shown below. Also, we have submitted a paper related to the inhibited mechanism by the polyamines to some journal. The latter paper is now in Revise.ArticleA.UEHARA,M.YASUKOCHI AND I.IMANAGA.Modulation of Ryanodine Binding to the Cardiac Ca^<2+> Release Channel by Arachidonic Acid. Journal of Molecular and Cellular Cardiology (1996) 28,43-51. Effects of arachidonic acid (AA) on the Ca^<2+> release channels in cardiac sarcoplasmic reticulum were examined by the ^3H-ryanodine binding method. The samples used were membrane vesicles of junction sarcoplasmic reticulum (JSR) and solubilized ryanodine receptor proteins. AA inhibited the amount of hot ryanodine bound to its receptor in both types of samples and this inhibitory effect was dose-dependent. The K_<hatf> values of the dose-response curve were 12 and 97 muM in the JSR membrane vesicles and the solubilized proteins, respectively. Moreover, Michaelis, Scatchard and Lineweaver-Burk analyzes were performed to evaluate K_<d+> B_<max> and K_d/B_<max> values. During exposure to AA,the K_d value increased while the B_<max> value decreased. These results suggest that AA directly modifies the structure of the ryanodine binding site.(c) 1996 Academic Press Limited

用平面脂质双层法、热配体结合法和钙离子外排法研究了生理活性物质对心肌肌浆网和线粒体单通道分子的影响<45>。对多种物质和离子通道进行了系统的筛选工作。这是最重要的发现，花生四烯酸和多胺的细胞质应用导致一个强有力的功能调节ryanodine受体通道。我们已经发表了一篇关于花生四烯酸及其类似物激活机制的论文，其摘要如下所示。另外，我们还向一些期刊提交了一篇关于多胺抑制机理的论文。后一篇论文现在发表在修订文章A.Uehara，M.Yasukochi和I.IMANAGA。花生四烯酸对Ryanodine结合心脏Ca^2+释放通道的调节。分子和细胞心脏病学杂志（1996）28，43 -51.用^3 H-兰尼碱结合法观察了花生四烯酸（AA）对心肌肌浆网Ca^2+释放通道的作用。使用的样品是肌浆网连接处（JSR）的膜囊泡和溶解的兰尼碱受体蛋白。AA抑制热ryanodine结合到其受体的量在这两种类型的样品，这种抑制作用是剂量依赖性的。剂量<hatf>-反应曲线的K值分别为12和97 μ M的JSR膜囊泡和溶解的蛋白质，分别。用米氏分析、Scatchard分析和Lineweaver-Burk分析计算了K_d+&gt; B_<max>和K_d/B_<max>值。在AA作用下，K_d值增加，而B<max>值下降。这些结果表明，AA直接修改的兰尼碱结合位点的结构。(c)1996年学术出版社

项目成果

Uehara, A., Yasukochi, M.and Imanaga, I.: "Modulation of ryanodine binding to the cardiac Ca^<2+> release channel by arachidonic acid." J.Mol.Cell.Cardiol. 28. 43-51 (1996)

Uehara, A.、Yasukochi, M. 和 Imanaga, I.：“通过花生四烯酸调节兰尼碱与心脏 Ca^2 释放通道的结合。”

Uehara,A.,Yasukochi,M.and Imanaga,I.: "Modulation of ryanodine binding to the cardiac Ca^<2+> release channel by arachidonic acid." J.Mol.Cell.Cardiol.28. 43-51 (1996)

Uehara,A.、Yasukochi,M. 和 Imanaga,I.：“通过花生四烯酸调节兰尼碱与心脏 Ca^2 释放通道的结合。”

Uehara,A.: "Calcium modulation of single SR potassium channel currents in heart muscle." J.Mol.Cell.Cardiol.25(In press). (1994)

Uehara,A.：“心肌中单个 SR 钾通道电流的钙调节。”