NEUROCHEMISTRY OF DRUG ABUSE

描述（申请人摘要）： 这是一个独立科学家奖的申请。长期 这项研究的目的是了解生物 神经递质多巴胺（DA）在几种结构中的意义 中枢神经系统在药物作用中的作用 虐待 计划进行两方面的调查。一个领域是审查 底物和转运蛋白的结构特征影响的动力学 人多巴胺和去甲肾上腺素转运蛋白，分别为hDAT和hNET。 大量的工作已经进入hDAT和hNET抑制剂的研究， 但是对衬底的特性研究较少。特别是， 关于胺基与邻苯二酚相互作用的假说 将测试转运蛋白中具有特定氨基酸的组。一 将使用最近开发的旋转圆盘电极伏安法 观察摄取和诱导外排的时间过程。的结果 这些研究将放在动力学研究的背景下， 印刷受体. 与上述研究相关的第二个领域是 hDAT和hNET底物的结合位点。其他人以前的工作 使用光亲和标记鉴定抑制剂结合的区域。的一部分 拟议中的工作是一项合作， 通过质谱法鉴定共价连接至所述蛋白质的特定氨基酸。 光亲和剂这项工作将通过使用新的 合成的光亲和底物以定位底物结合。

DESCRIPTION (Applicant's Abstract): This is a request for an Independent Scientist Award. The long term objective of this research is the understanding of the biological significance of the neurotransmitter dopamine (DA) in several structures of the central nervous system with respect to its role in the action of drugs of abuse. Two areas of investigation are planned. One area is the examination of the structural features of substrates and transporter affecting kinetics of the human dopamine and norepinephrine transporters, hDAT and hNET, respectively. A great deal of work has gone into the study of inhibitors of hDAT and hNET, but features of substrates have been less studied. In particular, hypotheses concerning the interaction of the amine group and the catechol groups with specific amino acids in the transporter will be tested. A recently developed method, rotating disk electrode voltammetry, will be used to observe the time course of uptake and induced efflux. The results of these studies will be put in the context of kinetic studies on a range of substrates. A second area, related to the above studies, is the localization of the binding site of substrates for hDAT and hNET. Previous work by others has identified regions of inhibitor binding using photoaffinity labels. Part of the proposed work is a collaboration to use recent progress in mass spectrometry to identify the specific amino acids covalently linked to the photoaffinity agents. This work will then be extended by using a newly synthesized photoaffinity substrate to localize substrate binding.