ON THE MECHANISM OF OXIDATIVE PHOSPHORYLATION

氧化磷酸化机制的探讨

• 批准号: 3268530
• 负责人: D R SANADI
• 金额: $ 27.32万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-07-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3268530
• 关键词: adenosinetriphosphatase; complementary DNA; cow; electron microscopy; electron transport; enzyme complex; enzyme linked immunosorbent assay; enzyme reconstitution; enzyme structure; gel filtration chromatography; genetic manipulation; heart metabolism; high performance liquid chromatography; membrane proteins; membrane reconstitution /synthesis; membrane structure; mitochondrial membrane; molecular cloning; monoclonal antibody; myocardium; nucleic acid sequence; oxidative phosphorylation; protein reconstitution; protein sequence; protein structure; radioimmunoassay; radiotracer; respiratory enzyme; respiratory protein

The research will focus on the terminal reactions of mitochondrial oxidative phosphorylation dealing with the utilization of the electrochemical proton gradient for the phosphorylation of ADP. The process is carried out by a vesicular H+-ATPase consisting of a hydrophobic membrane-embedded segment (Fo) and a hydrophilic, extrinsic multiprotein complex, F1-ATPase. Among the proteins of Fo is a 15,000 dalton, dithiol protein (coupling factor B or FB) which is essential for vectorial H+ translocation in Fo-F1. In its absence or when it is inhibited, the H+ release appears to be scalar, and ATP synthesis and the reverse H+ pumping reaction is blocked. Thus, detailed knowledge of the properties of FB and its mechanism of action are of crucial importance to a full understanding of one of the last unresolved major problems in metabolism. The aims include determination of the primary structure of FB by the cDNA cloning method, identification of proteins homologous with FB in the H+-ATPase preparations of other species, identification of the nearest neighbor proteins of FB, identification of another thiol protein involved in H+ conduction in Fo, and study of the mechanism of H+ conduction in Fo.

本研究将重点关注线粒体的终末反应， 氧化磷酸化处理的利用， 电化学质子梯度用于ADP的磷酸化。 的 这一过程是通过一个囊泡H +-ATP酶组成的疏水性 膜包埋片段（Fo）和亲水性外源性多蛋白 复合物，F1-ATP酶。 在Fo的蛋白质中，有一个15，000道尔顿的二硫醇 载体H+所必需的蛋白质（偶联因子B或FB） Fo-F1易位。 当它不存在或被抑制时， 释放似乎是标量，ATP合成和反向H+泵 反应受阻。 因此，详细了解FB的性质， 其作用机制对于充分了解 新陈代谢中最后未解决的主要问题之一。 目的包括通过cDNA序列测定FB的一级结构， 克隆方法、FB同源蛋白的鉴定 其他物种的H +-ATP酶制剂，最近的鉴定 FB的邻近蛋白，鉴定另一个参与的巯基蛋白 H~+在Fo中的传导，并探讨了H~+在Fo中的传导机制。