BIO-AND IMMUNO-CHEMICAL STUDIES ON OPIATE RECEPTORS

阿片受体的生物和免疫化学研究

• 批准号: 3211718
• 负责人: JAY A GLASEL
• 金额: $ 11.73万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1992-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3211718
• 关键词: antiidiotype antibody; chemical binding; conformation; gene expression; genetic recombination; histochemistry /cytochemistry; immunochemistry; laboratory mouse; laboratory rat; neuropeptide receptor; protein sequence

The purpose of this research is to combine the expertise of two laboratories in fundamental studies on the biochemistry and molecular biology of opiate receptors. Our laboratory has developed an immunochemical approach towards characterizing the receptors. In particular, this has involved the production of anti-idiotypic antibodies which competitively inhibit drug binding to the receptors. The other laboratory has been one of the world leaders in developing solubilization methods which are applicable to these particular membrane bound receptors. Anti-idiotypic antibodies which bind specifically to opiate receptors are to be used as probes in biochemical, molecular biological and immunocytological studies on opiate receptors. In the biochemical work the antibodies will be used in conjunction with reversible protein-protein cross-linking agents in order to solublize the entire receptor complexes. These will then be purified using immunoaffinity techniques in order to obtain receptor complexes which may be studied both in their entirtey as well as broken into their components for detailed characterization. In molecular biological work the anti-idiotypic antibodies will be used as probes for translational products in recombinant DNA experiments. With these techniques opiate receptor proteins will be sequenced indirectly. This information will be further used, in a second stage of the work, to sequence the genomic arrangements which give rise to the receptors. The immunocytological work will be directed towards exploring methods directed towards the end of fractionating opiate receptor-bearing neurons from the general population of central nervous tissue neurons. This latter, combined with the molecular biological information will ultimately lead to studies of; the control of opiate receptor expression in cells, the primary events following ligand recognition by the receptors, and the roles of receptors in pain path-ways and addictive liability of drugs.

这项研究的目的是将两者的专业知识结合起来 生物化学和生物化学基础研究实验室 阿片受体的分子生物学。我们的实验室有 开发了一种免疫化学方法来表征 感受器。特别是，这涉及到生产 竞争性抑制药物结合的抗独特型抗体 到感受器。另一个实验室是世界上最大的实验室之一 领导开发适用的增溶方法 这些特定的膜结合受体。 与阿片类药物特异性结合的抗独特型抗体 受体被用作生物化学、分子生物学中的探针 阿片受体的生物学和免疫细胞学研究。在……里面 抗体的生化工作将结合使用 使用可逆的蛋白质-蛋白质交联剂 溶解整个受体复合体。然后，这些将是 使用免疫亲和技术进行纯化，以获得 可作为整体研究的受体复合物 以及分解成它们的组件，以获得详细的 人物刻画。在分子生物学工作中，反独特型 抗体将被用作翻译产物的探针 重组DNA实验。使用这些技术可以使人产生鸦片 受体蛋白将被间接测序。此信息 将在工作的第二阶段进一步用于排序 产生受体的基因组排列。这个 免疫细胞学的工作将指向探索 针对分馏阿片类药物终结的方法 中枢神经系统一般人群中的受体神经元 神经组织神经元。后者与分子相结合 生物信息最终将导致对 阿片受体在细胞中表达的控制，主要事件 在受体识别配体之后，以及受体的作用 痛觉通路受体与药物成瘾易感性。