BIOCHEMISTRY AND GENETICS OF MEMBRANE FUNCTIONS

拟议研究的长期目标是提供更好的 了解结合蛋白依赖的运输系统。 我实验室可重组胶订的最新进展 蛋白质依赖的谷氨酰胺转运在分离的右侧-外-外 大肠杆菌的囊泡应该有助于结合的研究。 用物理化学方法研究蛋白质-膜相互作用。 谷氨酰胺在囊泡系统中转运的能量研究 得出了重要的发现，即三磷酸腺苷对 为运输提供所需的能源；继续 在这里提出的研究很可能导致最终的识别 难以捉摸的能量供体以及对其 生物合成。谷氨酰胺转运系统GLNP一直是 用重组DNA技术克隆和分析 已识别编码产品。遗传学和分子遗传学 我们建议进行实验，以研究 GLNP突变--有害效应的生理基础 GlnM蛋白和氮的过度生产- GLNP制度的监管。具体目标是：确定 右侧谷氨酰胺运输系统的能量供给者- 使用高标记~3H-的侧向和内向外的囊泡系统 ADP和~(32)P-PEP作为示踪剂；分离、纯化和鉴定 研究谷氨酰胺转运蛋白的遗传调控。 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；研究其生理生化 “杀手”基因glnM的作用研究 谷氨酰胺转运的功能方面。

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.