TRANSPORT ENZYMES IN SYNAPTIC VESICLES

突触小泡中的转运酶

• 批准号: 3395918
• 负责人: STANLEY MONROE PARSONS
• 金额: $ 17.71万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-03-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3395918
• 关键词: Chondrichthyes; acetylcholine; active transport; adenosinetriphosphatase; binding proteins; density gradient ultracentrifugation; drug receptors; electron microscopy; gel electrophoresis; high performance liquid chromatography; hydrolysis; ion exchange chromatography; ionophores; membrane permeability; neural information processing; neurochemistry; parasympathetic nervous system; radioassay; radiotracer; saltwater environment; spectrometry; synaptic vesicles; transport proteins

This project will elucidate the biochemical mechanisms responsible for storage of the neurotransmitter acetylcholine (ACh) by synaptic vesicles of nerve terminals. The guiding hypothesis is that recycling metabolically active VP2 vesicles carry out the majority of ACh active transport. A method for purifying VP2 vesicles from Torpedo californica electric organ will be developed. The gross physical and chemical properties of VP2 will be compared with those of the well-studied resting VP1 vesicles, and mechanistic aspects of the ACh storage process will be reinvestigated in VP2 vesicles. The VP2 vesicle polypeptides containing the ACh and drug inhibition binding sites of the ACh transport system will be identified by radioactive affinity labeling experiments. The proton pump ATPase which drives ACh storage, drug receptor and ACh transporter proteins will be purified in native detergent solubilized form. The isolated proteins will be reconstituted into liposomes and the ionic mechanism for each wll be characterized. Biochemical properties of each protein will be compared for the forms isolated from VP1 and VP2 vesicles in an effort to identify and localize regulation in the ACh storage system. It is expected that the project will develop the necessary background for biochemical study of ACh storage in mammalian brain and other organs. This should allow development of pharmacologic or other intervention strategies which would effect stimulation of ACh storage in and release from diseased or intoxicated central or peripheral cholinergic nerve terminals. A long-term goal is to find ameliorative strategies of use in the large number of neural dysfunctions, such as Alzheimer's disease, which have been linked to the cholinergic nerve terminal.

该项目将阐明负责的生化机制 突触小泡储存神经递质乙酰胆碱（ACh） 神经末梢。 指导性假设是代谢循环 活性 VP2 囊泡执行大部分 ACh 主动转运。 一个 一种加州鱼雷电器官VP2囊泡的纯化方法 将被开发。 VP2 的总体物理和化学性质将 与经过充分研究的静息 VP1 囊泡进行比较，并且 乙酰胆碱储存过程的机械方面将被重新研究 VP2 囊泡。 含有ACh和药物的VP2囊泡多肽 ACh 转运系统的抑制结合位点将通过以下方式确定 放射性亲和标记实验。 质子泵 ATP 酶 驱动乙酰胆碱储存、药物受体和乙酰胆碱转运蛋白 以天然洗涤剂溶解形式纯化。 分离出的蛋白质将 被重组为脂质体，并且每种的离子机制将是 特点。 将比较每种蛋白质的生化特性 从 VP1 和 VP2 囊泡中分离出的形式，以努力识别和 对乙酰胆碱存储系统进行本地化调节。 预计该项目将为以下领域发展必要的背景： 哺乳动物大脑和其他器官中乙酰胆碱储存的生化研究。 这 应允许制定药理学或其他干预策略 这会影响疾病中乙酰胆碱的储存和释放 或中毒的中枢或外周胆碱能神经末梢。 一个 长期目标是找到大范围使用的改进策略 一些神经功能障碍，例如阿尔茨海默氏病， 与胆碱能神经末梢相连。