MUTATIONAL LIBRARIES TO STUDY OPIOID RECEPTOR FUNCTION

描述：（申请人摘要） 点突变已被证明是强大的鉴定， 参与G蛋白偶联受体功能的结构氨基酸。 然而，产生适当的突变体需要预先考虑的 关于哪些氨基酸可能相关的假设。 由于多种 受体的功能，很难假定先验的参与， 特定的氨基酸以及氨基酸的特性 重要. 该建议旨在生成区域选择性随机 使用PCR策略的μ-阿片受体的突变文库。 最初，这些文库将用于筛选干扰 μ-阿片受体隔离。 以前我们已经发现， 阿片肽和生物碱的子集可以诱导 μ-阿片受体，而吗啡和许多临床使用的阿片类药物 不触发受体内化，或仅微弱地触发受体内化。 这 表明对人的免疫系统有类似作用的激动剂配体， 受体介导的信号传导可以对 受体运输 这项研究的目的是使用随机突变 库来了解配体依赖性受体的分子基础 内化 今后，这些结果应有助于澄清 内化在阿片类药物诱导的神经元损伤中的意义 宽容和依赖。

DESCRIPTION: (Applicant's Abstract) Point mutations have proven to be powerful for the identification of structural amino acids involved in Gprotein coupled receptor function. However, the generation of appropriate mutants necessitates a preconceived hypothesis regarding which amino acids may be relevant. For a number of receptor functions it is difficult to assume a priori the involvement of particular amino acids and what properties of the amino acids might be important. This proposal aims to generate region-selective random mutational libraries of the mu-opioid receptor using PCR strategies. Initially these libraries will be used to screen for mutations that perturb mu-opioid receptor sequestration. Previously we have found that endogenous opioid peptides and a subset of alkaloids can induce sequestration of the mu-opioid receptor, whereas morphine and many clinically-used opiate drugs do not trigger receptor internalization, or do so only weakly. This suggests that agonist ligands which have similar effects on receptor-mediated signaling can have dramatically different effects on receptor trafficking. The aim of the study is to use the random mutational libraries to understand the molecular basis for ligand-dependent receptor internalization. In the future, the results should help to clarify the significance of internalization in the development of opiate drug-induced tolerance and dependence.