THE THYLAKOID ENERGY TRANSDUCING ATPASE COMPLEX

类囊体能量转导ATP酶复合物

• 批准号: 3279371
• 负责人: BRUCE R SELMAN
• 金额: $ 11.65万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-08-01 至 1986-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3279371
• 关键词: Chlorophyta; adenine nucleotides; adenosine triphosphate; adenosinetriphosphatase; cell membrane; chloroplasts; circular dichroism; electron transport; enzyme substrate; fluorescence polarization; maleimides; molecular site; oxidative phosphorylation; phenazines; photosynthesis; plants; solvents; stereoisomer

In this proposal, we plan to investigate the energy transducing ATPase (CF0-CF1) complex associated with the thylakoid membranes of higher plants and the green algae Chlamydomonas reinhardi. Part I: Mechanistic problems related to the higher plant CF0-CF1 complex. Section A: Photoaffinity labels (2-azido adenine nucleotides) will be used to locate and identify the nucleotide regulatory binding site on membrane-bound CF1. Section B: (i) Distereomers of nucleoside phosphothioates will be used to determine the absolute stereochemistry of the ATPase reaction catalyzed by soluble CF1. (ii) Isotope trapping experiments will be performed to determine the kinetic mechanism of ATP synthesis and the binding constant for the first substrate to the ATP synthase active site. Section C: We will investigate the reversibility of the membrane-bound CF1 ATPase. Part A: We will investigate the mechanism of the reversible solvent-induced activation of the soluble ATPase using both physical and chemical methods. Part B: We will isolate and purify the CF0-CF1 complex from C. reinhardi. Part C: We plan to investigate the biochemical basis for the inability of isolated thylakoid membranes from C. reinhardi strain 2137 to catalyze photophosphorylation when cultured in the dark. Part D: We will measure the turnover of membrane-bound CF1 and the pool sizes of its subunits. SECTION D: We plan to isolate mutants of C. reinhardi defective in CF1 and characterize the resultant polypeptides.

在本提案中，我们计划研究能量转导 ATP 酶 (CF0-CF1) 与高等植物类囊体膜相关的复合物 和绿藻莱茵衣藻。 第一部分：机制问题 与高等植物 CF0-CF1 复合体有关。 A 部分：光亲和力 标签（2-叠氮基腺嘌呤核苷酸）将用于定位和识别 膜结合 CF1 上的核苷酸调节结合位点。 B 部分： (i) 核苷硫代磷酸酯的二对映异构体将用于测定 可溶性物质催化的 ATPase 反应的绝对立体化学 CF1。 (ii) 将进行同位素捕获实验以确定 ATP合成的动力学机制和第一个结合常数 ATP合酶活性位点的底物。 C 部分：我们将进行调查 膜结合 CF1 ATP 酶的可逆性。 A 部分：我们会 研究溶剂诱导的可逆活化机制 采用物理和化学方法制备可溶性 ATP 酶。 B部分：我们 将从莱茵衣藻中分离和纯化 CF0-CF1 复合物。 C 部分：我们 计划调查无法分离的生化基础 来自莱茵衣藻菌株 2137 的类囊体膜催化 在黑暗中培养时发生光磷酸化。 D 部分：我们将测量 膜结合 CF1 的周转及其亚基池大小。 D 部分：我们计划分离 CF1 和莱茵衣藻缺陷突变体 表征所得多肽。