BIOCHEMISTRY AND GENETICS OF MEMBRANE FUNCTIONS

膜功能的生物化学和遗传学

• 批准号: 3277552
• 负责人: JEN SHIANG HONG
• 金额: $ 21.6万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-12-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3277552
• 关键词: Escherichia coli; active transport; binding proteins; genetic mapping; glutamine; laboratory rabbit; membrane activity; membrane permeability; membrane structure; microorganism metabolism; molecular cloning; molecular genetics; mutant; nuclear magnetic resonance spectroscopy; nucleic acid sequence; plasmids; protein reconstitution; protein sequence; thin layer chromatography; transport proteins

The long-tem goals of the proposed research are to provide better understanding of the binding protein-dependent transport systems. Recent development in my laboratory of a reconstitutable binding protein-dependent glutamine transport in isolated right-side-out- vesicles of E. coli should facilitate the study of the binding protein-membrane interaction by physico-chemical methods. Energetic study of glutamine transport in the vesicle system has yielded the significant finding that ATP is essential for the generation of the needed energy donor for transport; continued study proposed here is likely to lead to the ultimate identification of the elusive energy donor as well as the elucidation of its biosynthesis. The glutamine transport systems glnP has been cloned and analyzed by recombinant DNA techniques and the encoded products identified. Genetic and molecular genetic experiments are proposed to study the physiological effects of the glnP mutations, the physiological basis of the deleterious effect of the overproduction of the glnM protein as well as the nitrogen- regulation of the glnP system. The specific aims are: to identify the energy donor of the glutamine transport system in the right- side-out and inside-out vesicle systems using highly labeled 3H- ADP and 32P-PEP as tracers; to isolate, purify and characterize glutamine transport proteins; to study the genetic regulation of the glutamine transport system; to investigate the metabolic and physiological effects of glutamine transport defects on glutamine and proline utilization; to study the physiological and biochemical effects of the "killer" gene glnM; to study the structural and functional aspects of glutamine transport.

拟议研究的长期目标是提供更好的 了解结合蛋白依赖性运输系统。 我的实验室最近开发了可重构绑定 分离的右侧向外的蛋白质依赖性谷氨酰胺转运 大肠杆菌的囊泡应该有助于结合的研究 通过物理化学方法研究蛋白质-膜相互作用。 囊泡系统中谷氨酰胺转运的能量研究 得出的重要发现是 ATP 对于 产生交通运输所需的能源；持续 这里提出的研究可能会导致最终的鉴定 难以捉摸的能量供体及其阐明 生物合成。 谷氨酰胺转运系统glnP 通过重组 DNA 技术进行克隆和分析 已识别的编码产品。 遗传和分子遗传学 建议进行实验来研究生理效应 glnP 突变，有害作用的生理基础 glnM 蛋白以及氮的过量产生 glnP 系统的调节。 具体目标是：确定 右谷氨酰胺转运系统的能量供体 使用高度标记的 3H- 的 side-out 和 inside-out 囊泡系统 ADP和32P-PEP作为示踪剂；分离、纯化和表征 谷氨酰胺转运蛋白；研究基因调控 谷氨酰胺转运系统；研究代谢和 谷氨酰胺转运缺陷对谷氨酰胺的生理影响 和脯氨酸的利用；研究生理和生化 “杀手”基因glnM的影响；研究结构和 谷氨酰胺转运的功能方面。