KINETICS AND MECHANISM OF CATECHOLAMINE TRANSPORTERS

描述：（申请人摘要） 多巴胺、去甲肾上腺素和5-羟色胺的转运蛋白是 滥用药物，如可卡因和安非他明，以及治疗药物 比如抗抑郁药 这些膜的结构和机理 跨越蛋白质仍然未知。 然而，最近的克隆和 这些转运蛋白的表达使得在更好地 了解这些蛋白质是如何工作的。 拟议的实验建立在 结合分子生物学的发展， 电化学测量技术的最新进展。 的动力学和 人去甲肾上腺素转运蛋白的机制将被检查， 快速伏安法 关于补充变更的假设 将测试底物结构和转运蛋白突变。 底物的氨基与氨基酸残基的相互作用 在跨膜区，一个人组成一组实验。 相互作用 跨膜区7中的残基的环羟基的折衷 第二组实验。 另一项实验旨在 表征被设计为用作 用于探测转运活性位点的光亲和标记。

DESCRIPTION: (Applicant's Abstract) The transporters for dopamine, norepinephrine, and serotonin are targets for drugs of abuse such as cocaine and amphetamine, and for therapeutic drugs such as the antidepressants. The structure and mechanism of these membrane spanning protein remains unknown. However, the recent cloning and expression of these transporters has allowed progress to be made in better understanding how these proteins work. The proposed experiments build on this recent progress by combining the developments in molecular biology with recent advances in electrochemical measurement technology. The kinetics and mechanism of the human norepinephrine transporter will be examined with rapid voltammetric methods. Hypotheses concerning complimentary alterations in substrate structure and mutation in the transporter will be tested. Interactions of the amino group of the substrate with an amino acid residue in transmembrane region one constitute one set of experiments. Interactions of the ring hydroxyls with residues in transmembrane region seven compromise a second set of experiments. An additional experiment is designed to characterize the transport characteristics of a substrate designed to act as photoaffinity label for probing the active site of transport.