Mechanisms Regulating Endocytosis of Opioid Receptors

阿片受体内吞作用的调节机制

• 批准号: 9318462
• 负责人: Mark E VonZastrow
• 金额: $ 35.3万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-28 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9318462
• 关键词: Acute; Adenylate Cyclase; Adrenergic Receptor; Affinity; Agonist; Alpha Cell; Arrestins; Behavior; Binding; Biological; Biological Assay; Biological Response Modifier Therapy; Biology; Biosensor; Brain Diseases; Catecholamine Receptors; Cell membrane; Cell model; Cell physiology; Cells; Cellular biology; Chemicals; Complement; Complex; Cyclic AMP; Data; Development; Drug Addiction; Drug Exposure; Drug Targeting; Drug effect disorder; Endocytosis; Endosomes; Enzymes; Event; Family; Family member; Funding; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-substrate; GTP-Binding Proteins; Generations; Goals; Heterotrimeric GTP-Binding Proteins; Human; Individual; Knowledge; Ligands; Link; Location; Mass Spectrum Analysis; Mediating; Memory; Methods; Mind; Molecular; Molecular Conformation; Molecular Target; Neurons; Opioid; Opioid Receptor; Pharmaceutical Preparations; Pharmacology; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Population; Process; Property; Protein Isoforms; Proteins; Public Health; Receptor Activation; Receptor Signaling; Recruitment Activity; Regulation; Research; Signal Transduction; Signaling Protein; Site; Specificity; System; Technology; Testing; Therapeutic; Translations; Work; addiction; base; beta-arrestin; desensitization; improved; in vivo; innovation; insight; link protein; mu opioid receptors; neural circuit; neurophysiology; new therapeutic target; programs; receptor; response; trafficking

PROJECT SUMMARY This proposal seeks to elucidate how G protein-coupled receptors (GPCRs) are regulated by naturally produced ligands and addictive drugs, focusing at the cell biological level as the critical bridge linking molecular and systems-level events and understanding. GPCRs represent the largest family of signaling receptors, comprise in aggregate the largest class of therapeutic drug targets, and mediate directly or indirectly the effects of all addictive drugs. Accordingly, while addictive drug action in the CNS is our particular focus, the proposed studies have broad potential application across GPCR family members and pathophysiological processes. Our over-arching goal is to develop a fundamental understanding of GPCR regulation, discovering the underlying cell biology and then elucidating its molecular basis, and through this path discover new targets and strategies for potential therapeutic manipulation of addictive and other complex brain disorders that are characterized by underlying disturbances of GPCR signaling or GPCR-dependent physiological regulation. Progress in the previous funding period focused on defining sites of regulatory phosphorylation in the mu opioid receptor, accomplishing this in intact human cells expressing the full spectrum of endogenous kinases at native levels. We defined two critical regions of phosphorylation and carried out detailed cell biological analysis of one of them, both in a heterologous cell model and a physiologically relevant population of CNS- derived neurons. During the course of these studies we made some major unanticipated progress, including development of conformational biosensor technology, discovery of GPCR signaling via heterotrimeric G proteins from endosomes, and discovery of an unprecedented behavior of arrestin proteins suggesting a new cellular operating mode of arrestins, downstream of and after dissociating from an activating GPCR. The proposed studies seek to develop and extend these fundamental new observations and develop them to the point of rational consideration as new molecular targets and cell-based strategies for therapeutics.

项目概要 该提案旨在阐明 G 蛋白偶联受体 (GPCR) 如何受到自然调节 生产配体和成瘾药物，重点关注细胞生物学水平，作为连接分子的关键桥梁 以及系统级事件和理解。 GPCR 代表最大的信号受体家族， 总体上包括最大类别的治疗药物靶点，并直接或间接介导 所有成瘾药物的影响。因此，虽然中枢神经系统的成瘾药物作用是我们特别关注的焦点， 拟议的研究在 GPCR 家族成员和病理生理学方面具有广泛的潜在应用 流程。我们的首要目标是对 GPCR 调控有一个基本的了解，发现 潜在的细胞生物学，然后阐明其分子基础，并通过这条路径发现新的靶点 以及对成瘾性和其他复杂脑部疾病的潜在治疗操纵策略 其特征是 GPCR 信号传导或 GPCR 依赖性生理调节的潜在紊乱。 上一个资助期的进展重点是确定 mu 中的调节性磷酸化位点 阿片受体，在表达全谱内源激酶的完整人类细胞中实现这一点 原生水平。我们定义了磷酸化的两个关键区域并进行了详细的细胞生物学研究 在异源细胞模型和中枢神经系统生理相关群体中对其中之一进行分析 派生神经元。在这些研究过程中，我们取得了一些意想不到的重大进展，包括 构象生物传感器技术的发展，通过异三聚体 G 发现 GPCR 信号传导 来自内体的蛋白质，以及抑制蛋白前所未有的行为的发现表明了一种新的 视紫红质抑制蛋白的细胞操作模式，在激活的 GPCR 下游和解离后。这 拟议的研究旨在发展和扩展这些基本的新观察结果，并将其发展到 作为新的分子靶点和基于细胞的治疗策略的理性考虑点。