NEUROCHEMISTRY OF DRUG ABUSE

描述(申请人摘要)： 这是对独立科学家奖的请求。从长远来看 本研究的目的是为了了解生物 神经递质多巴胺在中枢神经系统几种结构中的意义 中枢神经系统在药物作用中的作用 滥用职权。 计划进行两个领域的调查。其中一个领域是对 底物和转运体的结构特征对酶动力学的影响 人类多巴胺和去甲肾上腺素转运蛋白，分别为HDAT和HNET。 HDAT和HNET的抑制剂的研究已经进行了大量的工作， 但对底物的特性研究较少。特别是， 关于胺基与邻苯二酚相互作用的假说 将对转运蛋白中含有特定氨基酸的基团进行测试。一个 将使用最近开发的方法--旋转圆盘电极伏安法 观察摄取和诱导外排的时间进程。结果是 这些研究将放在一系列动力学研究的范围内 底物。 与上述研究相关的第二个领域是本地化 HDAT和hNET底物的结合部位。其他人之前的工作已经 使用光亲和标记识别抑制物结合的区域。部分 拟议的工作是一项合作，利用最新的大规模进展 光谱分析以确定特定氨基酸的共价连接 光亲和剂。这项工作将通过使用新的 合成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.