Structure and Function of the Yeast ATPase

描述(申请人提供)：线粒体ATP合成酶负责真核细胞在有氧条件下合成90%以上的细胞ATP。ATP是细胞用于几乎所有需要能量的过程的能量货币，如肌肉收缩(如心脏)和生物合成反应。这是对该酶进行深入研究的主要推动力。但最近的研究揭示了研究ATP合成酶的更大原因。三磷酸腺苷合成酶参与了许多与三磷酸腺苷合成活性相关或无关的其他关键过程。ATP合成酶是一个潜在的靶点，通过诱导细胞凋亡或调节血管生成来治疗癌症，治疗狼疮，通过增加代谢率从而提供热量限制或通过调节已知的参与延长寿命的受体来延长人类的寿命，预防心脏病，治疗眼病，如青光眼或黄斑变性，治疗肥胖，以及治疗细菌感染，如结核分枝杆菌或机会细菌铜绿假单胞菌引起的感染。这个项目的直接目标是了解ATP合成酶的结构/功能关系，并确定ATP合成酶的关键结构区域，这些区域如果被调节，就会使酶失活或受损。长期目标是针对这些地区进行合理的药物设计，以确定新的抗生素或药物。这个项目有两个目标。第一个目标是识别和了解分子结构特征，这些特征对于质子流与线粒体ATP合成酶合成ATP的耦合至关重要。第二个目标是确定ATP合成酶中的潜在靶点，用于基于Rational的药物设计。这些目标将通过相同的一系列实验来实现。具体地说，将在伽马亚基内或与其相关的区域被识别，这些区域具有严格的结构要求，如果受到干扰，会降低ATP合成酶的偶联效率。这种分析将使用基于结构的突变研究，然后对突变蛋白质进行生化和生物物理分析，并通过确定ATP合成酶的结构来完成。未来的研究将以这些区域为目标来开发抗生素，抗生素要么扰乱偶联效率，要么阻止病原菌的ATP合成酶伽马亚单位的旋转。

DESCRIPTION (provided by applicant): The mitochondrial ATP synthase is responsible for the synthesis of more than 90% of cellular ATP in the eukaryotic cell under aerobic conditions. ATP is the energy currency that the cell uses for nearly all energy requiring processes such as muscle contraction (as in the heart) and biosynthetic reactions. This has been a major impetus for the intense studies on this enzyme. But recent studies have revealed even greater cause to study the ATP synthase. The ATP synthase has been implicated in a number of other critical processes that are either related or unrelated to the activity of synthesis of ATP. The ATP synthase is a potential target to cure cancer either by eliciting apoptosis or by modulating angiogenesis, to treat Lupus, to extend the life- time of humans either by increasing the metabolic rate thereby providing caloric restriction or by modulating known receptors involved in extended life-span, to prevent heart disease, to treat eye disease, such as glaucoma or macular degeneration, to treat obesity, and to treat bacterial infections such as those caused by Mycobacterium tuberculosis or the opportunistic bacterium, Pseudomonas aeruginosa. The immediate goal of this project is to understand the structure/function relationship of the ATP synthase and to identify critical structural regions of the ATP synthase, which if modulated, inactivate or impair the enzyme. The long-term goal is to target these regions for rational drug design to identify new antibiotics or drugs. There are two objectives for this project. The first objective is to identify and understand molecular structural features critical for the coupling of the flow of protons with the synthesis of ATP by the mitochondrial ATP synthase. The second objective is to identify potential targets in the ATP synthase for rational based drug design. These objectives will be obtained with the same set of experiments. Specifically, regions will be identified within, or associated with, the gamma-subunit, which have strict structural requirements and if perturbed, reduces the efficiency of the coupling of the ATP synthase. This analysis will be done using structure-based mutagenesis studies followed by biochemical and biophysical analysis of the mutant proteins and by structure determination of the ATP synthase. Future studies will target these regions for the development of antibiotics, which either perturb the coupling efficiency or block rotation of the gamma-subunit of the ATP synthase from pathogenic bacteria.