SYNTHESIS OF CONSTITUTIVELY ACTIVE SEROTONIN RECEPTORS

拟议研究的主要目标是探索结构- 5-羟色胺1型(5HT1)受体的功能关系 激动剂结合部位。为实现这一目标，将努力 通过以下方式明确识别神经递质结合部位 结构性地激活5-羟色胺结合部位的受体。 这项研究的最终目标是合理地使用这些 活性受体研究5HT1受体的长期效应 激活。这项提案专门针对 非天然氨基酸在5HT1A受体中的掺入及其作用 在完整的细胞中表达。以下是研究报告 目标： L。设计和合成非天然氨基酸，包括 它们侧链中的5-羟色胺本身。 2.将这些非天然氨基酸加入到推定的 利用无义密码子抑制5HT1a激动剂结合位点 非天然氨基酸掺入非洲爪哇卵母细胞的方法。 3.功能性5HT1A受体的鉴定 功能偶联、共表达的电生理分析 鸟嘌呤结合蛋白(G蛋白)激活，向内整流 钾(K+)通道。

The primary goal of the proposed research is to probe structure- function relationships within the serotonin type 1 (5HT1) receptor agonist binding site. To achieve this goal, efforts will be made to unambiguously identify the neurotransmitter binding site by constitutively activating the receptor at the serotonin binding site. The ultimate goal of this research is to use these constitutively active receptors to study the long-term effects of 5HT1 receptor activation. This proposal is specifically focused on the incorporation of unnatural amino acids into 5HT1A receptors and their expression in intact cells. The following are the research objectives: l. Design and synthesis of unnatural amino acids that include serotonin itself in their side chains. 2. Incorporation of these unnatural amino acids into the putative 5HT1A agonist binding site using the nonsense codon suppression method for unnatural amino acid incorporation in Xenopus oocytes. 3. Identification of functional 5HT1A receptors through electrophysiological analysis of functionally coupled, coexpressed guanine binding protein (G-protein) activated, inwardly rectifying potassium (K+) channels.