Structural Basis of Opioid Receptor Function

阿片受体功能的结构基础

基本信息

批准号：
8590733
负责人：
Brian K Kobilka
金额：
$ 48.25万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-15 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8590733
关键词：
Acute Pain Address Adverse effects Agonist Architecture Behavior Binding Binding Proteins Biochemical Buprenorphine Cause of Death Chemicals Codeine Complex Coupling Crystallography Development Electron Microscopy Electron Spin Resonance Spectroscopy Europe Exhibits Family member Fluorescence Spectroscopy G-Protein-Coupled Receptors G-substrate GTP-Binding Proteins Goals Heroin Immunoglobulin Fragments Leucine Enkephalin Ligand Binding Ligands Mediating Methadone Molecular Conformation Morphinans Morphine NMR Spectroscopy North America Opioid Opioid Receptor Opium Overdose Pain Peptides Pharmaceutical Preparations Pharmacology Preparation Process Property Proteins Receptor Activation Resolution Signal Transduction Structure Techniques Therapeutic Translating base chronic pain delta opioid receptor desensitization design drug efficacy experience insight member mu opioid receptors nanobodies particle preference public health relevance receptor receptor coupling receptor function small molecule three dimensional structure tool

Acute Pain Address Adverse effects Agonist Architecture Behavior Binding Binding Proteins Biochemical Buprenorphine Cause of Death Chemicals Codeine Complex Coupling Crystallography Development Electron Microscopy Electron Spin Resonance Spectroscopy Europe Exhibits Family member Fluorescence Spectroscopy G-Protein-Coupled Receptors G-substrate GTP-Binding Proteins Goals Heroin Immunoglobulin Fragments Leucine Enkephalin Ligand Binding Ligands Mediating Methadone Molecular Conformation Morphinans Morphine NMR Spectroscopy North America Opioid Opioid Receptor Opium Overdose Pain Peptides Pharmaceutical Preparations Pharmacology Preparation Process Property Proteins Receptor Activation Resolution Signal Transduction Structure Techniques Therapeutic Translating base chronic pain delta opioid receptor desensitization design drug efficacy experience insight member mu opioid receptors nanobodies particle preference public health relevance receptor receptor coupling receptor function small molecule three dimensional structure tool

展开

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to determine the structural basis for opioid receptor pharmacology and function. Opioid receptors constitute the major and the most effective target for the treatment of pain. The use of opioid drugs acting at these receptors is however a leading cause of death by overdose in Europe and North America. Both beneficial and adverse effects of illicit opioid drugs (opium, heroin) as well as approved therapeutics (morphine and codeine) are mediated by the activation of mu opioid receptor (¿-OR), a member of the G protein-coupled receptors (GPCRs) superfamily. To understand the structural basis for ¿-OR function, we obtained the first high-resolution crystal structure of an inactive state of this receptor along with a closely relate family member the delta-opioid receptor (¿-OR). These structures provide new insights into OR preference for specific antagonist drugs but do not address important questions regarding OR activation mechanisms and, in particular, opioid drug efficacy. We therefore propose to characterize the structural basis of opioid receptor activation using a combination of biochemical and biophysical approaches. Specific Aims include: Aim 1. Determine active state structures of ORs The inactive state structures of the ¿-OR and ¿-OR provided the first structural insights into the binding mode of morphinan antagonists. The goal of this aim is to obtain structural insights into the process of opioid receptor activation of the G protein Gi. We will initially focus on the use of crystallography and single particle electron microscopy to obtain three-dimensional structures of the ¿-OR-Gi and ¿-OR-Gi complexes. We will also develop a panel of camelid antibody fragments (nanobodies) that stabilized ligand-specific conformational states for crystallography. These will be used to determine the structural basis for the different functional properties of opioid receptor agonists. Aim 2. Conformational dynamics of OR structure and activation. OR activation and more generally GPCR activation involves a complex allosteric coupling between ligand binding and G protein coupling domains that is poorly understood. Crystal structures offer a limited number of static snapshots of this dynamic process. We therefore propose to develop and apply biophysical approaches to characterize the structural plasticity and dynamic properties of ORs and to determine how this is translated into signaling complexity and ligand efficacy. Inactive state crystal structures will constitute an important starting point for designing and interpreting biochemical and biophysical studies, some of which include fluorescence, EPR and NMR spectroscopy. These studies will provide new insights into opioid ligand efficacy and the differences between small molecule and peptide ligands.

描述（由适用提供）：该提案的目的是确定阿片受体药理学和功能的结构基础。阿片受体构成了治疗疼痛的主要和最有效的靶标。但是，在这些受体上作用的阿片类药物的使用是欧洲和北美过量的主要死亡原因。非法阿片类药物（鸦片，海洛因）以及批准的治疗（吗啡和可待因）的有益和不利影响均由G蛋白偶联受体的成员激活MU阿片受体（�-OR）介导（GPCR）超家族。为了理解 - 或功能的结构基础，我们获得了该受体不活跃状态的第一个高分辨率晶体结构，以及密切相关的家族成员Delta-Apoile受体（ - 或）。这些结构为特定拮抗剂药物提供了新的见解或偏爱，但并未解决有关或激活机制，尤其是阿片类药物效率的重要问题。因此，我们建议使用生化和生物物理方法的组合来表征阿片受体激活的结构基础。具体目的包括：目标1。确定ORS的活性状态结构，即“或�”和“或€”或提供了第一个结构性见解，以对形态拮抗剂的结合模式。该目标的目的是获得对G蛋白GI的阿片受体激活过程的结构见解。我们最初将专注于晶体学和单个颗粒电子显微镜的使用，以获得 - 或GI和 - gi和 - 或GI复合物的三维结构。我们还将开发一个骆驼抗体片段（Nanobote）稳定了特定配体会议状态的晶体学状态。这些将用于确定阿片受体不同功能特性的结构基础。目标2。结构和激活的构象动力学。或激活和更普遍的GPCR激活涉及配体结合和G蛋白偶联域之间的复杂变构耦合，这些耦合域知之甚少。晶体结构提供了这种动态过程的有限数量的静态快照。因此，我们建议开发和应用生物物理方法来表征ORS的结构可塑性和动态特性，并确定如何将其转化为信号复杂性和配体效率。非活性状态晶体结构将构成设计和解释生化和生物物理研究的重要起点，其中一些包括荧光，EPR和NMR光谱。这些研究将为阿片类配体效率以及小分子和胡椒配体之间的差异提供新的见解。