BIOCHEMISTRY AND GENETICS OF MEMBRANE FUNCTIONS

拟议研究的长期目标是提供更好的 结合蛋白依赖的运输系统的理解。 最近在我的实验室的一个可重构的绑定的发展 蛋白质依赖的谷氨酰胺转运在分离的右侧出， E.大肠杆菌中的蛋白质应该有助于研究 蛋白质-膜相互作用的物理化学方法。 谷氨酰胺在囊泡系统中转运的研究 得出了重要的发现，ATP是必不可少的， 产生运输所需的能源捐助者;续 这里提出的研究可能会导致最终确定 的难以捉摸的能源捐助者，以及阐明其 生物合成 谷氨酰胺转运系统Glutamate已被 通过重组DNA技术克隆和分析， 已识别的编码产品。 遗传和分子遗传学 实验提出了研究的生理效应， glycoprotein突变，有害作用的生理基础， glnM蛋白和氮的过量生产 调节glomerate系统。 具体目标是： 谷氨酰胺转运系统的能量供体， 侧出和内面外囊泡系统使用高度标记的3 H- ADP和32 P-PEP作为示踪剂;分离、纯化和表征 谷氨酰胺转运蛋白;研究遗传调控 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效应 脯氨酸利用率的研究; “杀手”基因glnM的影响;研究结构和 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.