PROTON TRANSLOCATION THROUGH F1F0 ATP SYNTHASE

三磷酸腺苷合成酶(F1F0)是三磷酸腺苷合成酶的中心酶 在几乎所有的生物体中进行能量传递。能源匮乏 转导导致一组已知的悲剧性遗传性人类疾病 统称为线粒体肌病。很可能没有那么多 能量新陈代谢方面的有害问题仍将不被发现 临床实验室的能量代谢问题将在#年仍未被发现 临床实验室在许多其他方面起到了促进作用 健康问题，如心脏病。能量代谢的改变是 在大多数快速生长的肿瘤中也发现了这种情况。F1F0三磷酸腺苷合成酶通过 利用质子在生物膜上转移的能量 推动ADP的磷酸化，使其产生ATP。的长期目标是 这项研究是为了通过对质子运动的了解 分子水平上的酶。分子遗传学和生物化学是 用作统一的方法。大肠杆菌F1F0三磷酸腺苷基因突变的研究 合酶是通过定点突变产生的，而合酶的作用 通过一系列生化分析研究突变。就这样， 质子酶中特定氨基酸的功能作用 传导性可以被评估。与学习相关的困难 通过传统生物化学方法获得的疏水蛋白质已经受挫。 试图获得质子转移域的结构模型 F1F0三磷酸腺苷合成酶。另一种使用分子生物学的方法来插入 化学反应的靶氨基酸进入膜结合部分 使用了这种酶。然后目标氨基酸的环境就可以 通过以前不能使用的既定方法进行探测。 最后，测试通过实验生成的假设的方法 高等生物体中的细菌F1F0ATP合成酶是 探索过了。具体的方法包括建造一种混合动力车 含有人类质子转运系统的细菌酶和 最近邻细菌质子转运系统的研究 哺乳动物线粒体酶的近亲，球形红杆菌。

Adenosine triphosphate synthase (F1F0 ATP synthase) is the central enzyme for energy transduction in virtually all organisms. Deficiencies in energy transduction cause a group of tragic inheritable human disorders known collectively as mitochondrial myopathies. It is likely that less deleterious problems in energy metabolism will remain undetected in the clinical laboratory problems in energy metabolism will remain undetected in the clinical laboratory while acting as a contributing factor in many other health difficulties such as heart disease. Altered energy metabolism is also found in most rapidly growing tumors. F1F0 ATP synthases function by utilizing the energy of proton translocation across biological membrane to drive the phosphorylation of ADP making ATP. The long-term objective of the research is to obtain an understanding of proton movement through the enzyme at the molecular level. Molecular genetics and biochemistry are used as a unified approach. Mutations in the Escherichia coli F1F0 ATP synthase are generated by site-directed mutagenesis and the effects of the mutations investigated by a battery of biochemical analyses. In this way, the functional roles of specific amino acids within the enzyme in proton conductivity can be assessed. Difficulties associated with studying hydrophobic proteins via traditional biochemical approaches has frustrated attempts to gain a structural model of the proton translocation domain in F1F0 ATP synthase. An alternate approach using molecular biology to insert chemically reactive target amino acids into the membrane-bound portion of the enzyme is employed. The environment of the target amino acid can then be probed by established methods which could not be used previously. Finally, methods for testing hypotheses generated by experimentation with the bacterial F1F0 ATP synthase in the enzymes of higher organisms are explored. Specific approaches include the construction of a hybrid bacterial enzyme containing the human proton translocation system and the investigation of the proton translocation system of the nearest bacterial relative of the mammalian mitochondrial enzyme, Rhodobacter sphaeroides.