TRANSPORT ENZYMES IN SYNAPTIC VESICLES

突触小泡中的转运酶

• 批准号: 3395920
• 负责人: STANLEY MONROE PARSONS
• 金额: $ 19.86万
• 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/3395920
• 关键词: 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; 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 通过突触囊泡的神经递质乙酰胆碱（ACH）的储存 神经终端。 指导假设是代谢回收 主动VP2囊泡进行大部分ACH主动转运。 一个 从Torpedo Californica电器器官纯化VP2囊泡的方法 将被开发。 VP2的物理和化学特性将 将其与精心研究的静止VP1囊泡和 ACH存储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传输系统的抑制结合位点将通过 放射性亲和力标记实验。 质子泵ATPase 驱动ACH存储，药物受体和ACH转运蛋白蛋白将是 以天然洗涤剂溶解形式纯化。 孤立的蛋白质将 被重构为脂质体和每个wll的离子机制 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，以识别和 在ACH存储系统中本地化调节。 预计该项目将为 哺乳动物脑和其他器官中ACH存储的生化研究。 这 应该允许制定药理学或其他干预策略 这将影响刺激ACH存储中的储存和从患病中释放 或陶醉的中央或周围胆碱能神经末端。 一个 长期目标是在大型中找到改善使用策略 神经功能障碍的数量，例如阿尔茨海默氏病 与胆碱能神经末端有关。