BIOENERGETIC ASPECTS OF ALCOHOL CONSUMPTION

本申请是对ADAMHA研究科学家的要求 发展奖/二级威廉S。Thayer博士 的 该奖项将使塞耶博士能够全身心投入研究， 并将提高他的科学能力， 拓宽他在酒精相关研究方面的经验， 使用新的实验技术。 后者将包括 免疫化学和核磁共振光谱。 这项拟议中的研究将研究乙醇和 乙醇消耗对线粒体膜组装和 线粒体生物能量学 长期喂食乙醇的大鼠 用作酒精性脂肪肝的动物模型。 分离 肝细胞，肝线粒体，磷酸化亚线粒体 颗粒和磷脂囊泡， 线粒体蛋白质，特别是细胞色素氧化酶，将被 用作实验系统。 细胞色素的一种非活性形式 氧化酶，最近通过免疫化学技术鉴定， 酒精大鼠的亚线粒体颗粒，将被分离 和表征了 血红素和蛋白质的生物周转 细胞色素氧化酶和其他线粒体 膜蛋白，在体内将确定从时间进程 在用特异性标记后放射性降低 前体 醇对聚乙烯醇合成和组装的影响 细胞色素氧化酶和其它线粒体膜蛋白， 将通过免疫化学技术使用分离的 肝细胞 线粒体膜蛋白的重构 磷脂囊泡将被研究作为一个模型系统， 膜组件 脂质成分和物理的作用 测定蛋白质含量和酶的性质 将探索重组囊泡的活性。 的影响 乙醇消耗对线粒体生物能量学和功能的影响 在肝细胞中，将通过检查调节 呼吸速率、电化学质子 梯度和腺嘌呤核苷酸磷酸化系统。 乙醇对质子转移反应的影响 电化学质子梯度的产生也将 研究了 这些研究将有助于了解 酒精消费和意志的生物能量后果 提供有关乙醇基本机制的见解 毒性

This application is a request for an ADAMHA Research Scientist Development Award/Level II for William S. Thayer, Ph.D. The award will allow Dr. Thayer to devote his full efforts to research, and will enhance his scientific capabilities by allowing him to broaden his experience in alcohol-related research and learn the use of new experimental techniques. The latter will include immunochemistry and nuclear magnetic resonance spectroscopy. The proposed research will study the effects of ethanol and ethanol consumption on mitochondrial membrane assembly and mitochondrial bioenergetics. Rats fed ethanol chronically will be used as an animal model for alcoholic fatty liver. Isolated hepatocytes, liver mitochondria, phosphorylating submitochondrial particles, and phospholipid vesicles reconstituted with specific mitochondrial proteins, particularly cytochrome oxidase, will be used as experimental systems. An inactive form of cytochrome oxidase, recently identified by immunochemical techniques in submitochondrial particles from alcoholic rats, will be isolated and characterized. The biological turnover of heme and protein components of cytochrome oxidase, and other mitochondrial membrane proteins, in vivo will be determined from time courses of decreases in radioactivity following labeling with specific precursors. Effects of alcohol on the synthesis and assembly of cytochrome oxidase, and other mitochondrial membrane proteins, will be investigated by immunochemical techniques using isolated hepatocytes. Reconstitution of mitochondrial membrane proteins into phospholipid vesicles will be studied as a model system for membrane assembly. The role of lipid composition and physical properties in determining the protein content and enzymatic activity of reconstituted vesicles will be explored. Effects of ethanol consumption on mitochondrial bioenergetics and function in the liver cell will be studied by examining the regulatory relationships between respiration rate, electrochemical proton gradient and the adenine nucleotide phosphorylation system. Effects of ethanol on proton translocation reactions involved in generation of the electrochemical proton gradient will also be investigated. These studies will contribute to an understanding of the bioenergetic consequences of alcohol consumption and will provide insights concerning fundamental mechanisms of ethanol toxicity.