STRUCTURE/FUNCTION OF THE MITOCHONDRIAL K-ATP CHANNEL

线粒体 K-ATP 通道的结构/功能

• 批准号: 2900894
• 负责人: Keith D Garlid
• 金额: $ 25.16万
• 依托单位: OREGON GRADUATE INSTITUTE SCIENCE & TECH
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2900894
• 关键词: Escherichia coli; Saccharomyces cerevisiae; adenosine triphosphate; electron transport; gene expression; genetic library; glyburide; laboratory rat; membrane activity; mitochondrial membrane; molecular cloning; northern blottings; potassium channel; protein structure function; receptor binding; site directed mutagenesis; sulfonylurea

The mitochondrial KATP channel (mitoKATP) regulates matrix volume. Matrix volume, in turn, is hypothesized to control the activation state of electron transport. MitoKATP opens and closes in response to cellular levels of ATP, GTP and long-chain acyl-CoA esters. It is also a pharmacological receptor for K+ channel openers and the antidiabetic sulfonylureas. The broad objectives of this project are (1) to understand the molecular mechanisms of regulated K+ flux through mitoKATP; and (2) to understand the metabolic signalling role of mitoKATP activity in E. coli and/or S. cerevisiae; (3) to elucidate the molecular regulatory mechanisms of native and mutagenized mitoKATP; and (4) to test hypotheses relating to the functional role of mitoKATP. Peptide sequence for the sulfonylurea receptor subunit will be obtained and used to isolate its cDNA. Development of an expression system will be followed by protein isolation, reconstitution and assay of regulated K+ flux. K+ flux, measured with a K+ -specific fluorescent probe, will be assayed for inhibition by ATP, palmitoyl CoA, and glyburide, and reversal of inhibition by GTP and K+ channel openers. K+ flux will also be measured in intact mitochondria in order to examine the physiological role of mitoKATP using control analysis.

线粒体KATP通道（Mitokatp）调节矩阵体积。 依次假设矩阵体积可以控制激活状态 电子传输。 Mitokatp响应于 ATP，GTP和长链酰基-COA酯的细胞水平。 也是 K+通道开启器和抗糖尿病的药理受体 磺酰脲。 该项目的广泛目标是（1） 了解调节的K+通量的分子机制 mitokatp; （2）了解 大肠杆菌和/或酿酒酵母中的mitokatp活性； （3）阐明 天然和诱变Mitokatp的分子调节机制；和 （4）测试与Mitokatp功能作用有关的假设。 将获得磺酰脲受体亚基的肽序列 并用于隔离其cDNA。 表达系统的开发将 然后进行蛋白质隔离，重建和调节的测定 K+通量。 用K+特异性荧光探针测量的K+通量将 通过ATP，Palmitoyl COA和Glyburide抑制，并进行测定 GTP和K+通道开启器的抑制作用逆转。 K+通量也将 在完整的线粒体中测量以检查生理 使用控制分析的MitokATP的作用。