SYNAPTIC VESICLE CATECHOLAMINE/AEROTINA

突触小泡儿茶酚胺/Aerotina

• 批准号: 3401362
• 负责人: JOSEPH A NEAR
• 金额: $ 8.88万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3401362
• 关键词: adenosine triphosphate; affinity chromatography; affinity labeling; biomarker; brain cell; catecholamine inhibitor; catecholamines; central nervous system; chemical association; chemical binding; chromaffin cells; cow; crosslink; electrochemistry; electrophoresis; glycoproteins; isoelectric point; molecular weight; neurochemistry; radioassay; reserpine; serotonin; synaptic vesicles; tetrabenazine; tritium

The accumulation of catecholamines and serotonin in synaptic vesicles of the CNS is mediated by a transport system (porter) located in the vesicle membrane. This system is the primary target for several pharmacological agents that have been used extensively in research and are of considerable clinical interest. The long- term objectives of the proposed research include identification, reconstitution and characterization of the molecular entities directly involved in transport. Of particular interest are the subunit structure of the porter, and localization and orientation of binding sites for inhibitors such as reserpine and tetrabenazine. These two agents appear to represent distinct classes of inhibitors that may interact with different subunits of the porter complex in order to achieve these goals photoaffinity analogs of tetrabenazine, reserpine and substrates will be used to identify relevant components of the complex ln synaptic vesicles from beef brain, and labeled components will be characterized with respect to molecular weight isoelectric point, and glycoprotein content by lectin affinity chromatography. The effect of membrane energization on photoaffinity labeling will also be assessed. The respective binding components will be solubilized and purified by conventional biochemical methods, including affinity chromatography using inhibitor binding or photoaffinity labeling to follow activity. The ability of purified components to bind substrates and inhibitors will be compared, and the pH dependence of such binding assessed. The type of inhibition exerted by various agents on dopamine transport and reserpine and dihydrotetrabenazine binding to purified components will be examined. In addition to a better understanding of the structure and organization of the porter, the proposed work should yield information useful in the design of new catecholamine depleting therapeutic agents with potential utility in clinical situations that require a reduction of catecholamine available for release to the synapse.

儿茶酚胺和5-羟色胺在突触中的蓄积 中枢神经系统的囊泡是由运输系统(Porter)调节的 位于囊泡膜中。这个系统是主要的目标 已被广泛使用的几种药理药物 在研究中，并具有相当大的临床价值。长的- 拟议研究的长期目标包括识别、 分子实体的重构与表征 直接参与交通运输。特别值得关注的是 搬运工的亚基结构，以及定位和定位 利血平和利血平等抑制剂的结合部位 四苯那嗪。这两个特工似乎代表着不同的 可能与不同亚基相互作用的几类抑制剂 以实现这些目标的光亲和性 四苯肼、利血平和底物的类似物将用于 辨别复合体突触小泡的相关成分 来自牛脑，标记的成分将被表征为 关于分子量等电点和糖蛋白 凝集素亲和层析法测定含量。膜的作用 还将评估对光亲和标记的激励作用。这个 相应的结合组分将通过以下方式溶解和纯化 常规生化方法，包括亲和层析 使用抑制剂结合或光亲和标记进行跟踪 活动。纯化成分与底物结合的能力 和抑制剂将进行比较，以及这些药物的pH依赖性 已评估约束性。不同药剂施加的抑制类型 多巴胺转运与利血平和二氢四苯嗪 将检查与纯化组件的结合情况。除了……之外 更好地了解组织的结构和组织 波特，拟议的工作应该产生有用的信息在 新型儿茶酚胺耗竭治疗药物的设计 在需要减少的临床情况下的潜在效用 可以释放到突触的儿茶酚胺。