PROTON TRANSLOCATION THROUGH F1F0 ATP SYNTHASE

三磷酸腺苷合成酶（F1F0 ATP合成酶）是该酶的中心酶 几乎所有生物体的能量转换。 能源赤字 转导导致一组悲剧性的遗传性人类疾病， 统称为线粒体肌病 很可能， 能量代谢中的有害问题将在 临床实验室在能量代谢方面的问题仍然未被发现， 临床实验室虽然在许多其他方面起着促进作用， 健康问题，如心脏病。 能量代谢的改变 也见于大多数快速生长的肿瘤。 F1F0 ATP酶的功能是 利用质子跨生物膜迁移的能量， 驱动ADP磷酸化生成ATP。 的长期目标 这项研究的目的是通过质子的运动来了解质子的运动。 分子水平上的酶。 分子遗传学和生物化学 作为一种统一的方法。 大肠杆菌F1F0 ATP中的突变 合成酶的作用是通过定点诱变产生的， 通过一系列生化分析研究突变。 通过这种方式， 质子中酶内特定氨基酸的功能作用 可以评估电导率。 与学习相关的困难 疏水蛋白通过传统的生物化学方法已经失败， 试图获得质子转运结构域的结构模型， F1F0 ATP合成酶。 另一种方法是利用分子生物学 化学反应性的目标氨基酸进入膜结合的部分， 使用酶。 然后，目标氨基酸的环境可以 可以用以前不能使用的既定方法进行探测。 最后，本文介绍了检验实验假设的方法， 细菌F1F0 ATP合酶在高等生物的酶中是 探讨了 具体做法包括建设一个混合 含有人质子易位系统的细菌酶和 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，类球红细菌。

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.