Structure and Function of CB2 Cannabinoid Receptor

CB2 大麻素受体的结构和功能

基本信息

批准号：
7440803
负责人：
ZHAO-HUI SONG
金额：
$ 4.04万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-30 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7440803
关键词：
Absence of pain sensation Adverse effects Affinity Affinity Chromatography Agonist Agreement Amino Acids Analgesics Arts Autoimmune Diseases Binding Biochemical Biogenic Amine Receptors CNR2 gene Cannabinoids Cellular biology Characteristics Chemicals Class Cloning Collaborations Condition Conserved Sequence Cysteine Decompression Sickness Development Disulfide Linkage Disulfides Dopamine D2 Receptor Ensure Ethers Ethyl Ether Exhibits Family Fingerprint Fluorescence Spectroscopy Future G-Protein-Coupled Receptors Glaucoma Goals Growth Helix (Snails)Hydrogen Bonding Hydroxyl Radical Immune Immune system Immunologic Receptors Inflammation Ions Ligand Binding Ligands Liquid Chromatography Maps Marijuana Mass Spectrum Analysis Medical Metals Methodology Methods Modeling Modification Molecular Molecular Biology Molecular Conformation Mutagenesis Mutation Nature Neuraxis Nuclear Magnetic Resonance Numbers Peptide Fragments Peptides Peripheral Pharmaceutical Preparations Pharmacology Pichia Positioning Attribute Post-Translational Protein Processing Procedures Process Production Proline Property Protein Chemistry Protein Overexpression Proteins Proteomics Purpose Receptor Activation Research Design Research Personnel Research Proposals Resolution Rhodopsin Running Site Source Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Structure Surface System Testing Therapeutic Therapeutic Agents Time Tissues Transmembrane Domain Work X-Ray Crystallography Yeasts base cannabinoid receptor design disulfide bond ear helix improved insight mimetics molecular modeling novel programs protein aminoacid sequence protein structure receptor receptor structure function scale up tool

项目摘要

The long-term goal of this research proposal is to identify the molecular basis underlying the functions of the CB2 cannabinoid receptor. To achieve this purpose, the following steps will be taken. In Aim 1 and 2, the interactive mutagenesis, molecular modeling and new compound testing studies will be carried out. This combined approach has advanced our understanding, and should continue to reveal new insights on how cannabinoid ligands are recognized by their receptors, the origin of receptor subtype-selectivity, and how _these receptors are activated/inactivated. Cannabinoid receptors share many of the conserved sequence motifs seen in other well-studied G protein-coupled receptors (GPCRs), such as rhodopsin and biogenic amine receptors. However, several helix-bending residues and motifs, and a key disulfide bridge present in many other GPCRs are missing in cannabinoid receptors. In Aim 3, Ligand binding crevice exposure of two transmembrane domains of CB2 cannabinoid receptor will be mapped using substituted-cysteine accessibility method. This approach should elucidate structural and functional consequences of the sequence divergence in these trasmembrane domains of cannabinoid receptors. In Aim 4, Disulfide bond formation of CB2 receptor will be determined directly by the state-of-art methodologies of mass spectrometry. Investigating the unique disulfide bond or absence of a disulfide bond should provide important helix folding information of CB2 receptor, and this will have important implications regarding ligand binding crevice and activation mechanisms of this receptor. Finally, in Aim 5, effort will be devoted to over-express and purify large amounts of functional CB2 receptor. This challenging task, if successfully completed, should have a significant impact by providing the badly needed pure receptor proteins for future high-resolution biophysical studies. Overall, This study should help us to understand in more molecular detail the structure and function of CB2 receptor. CB2 is primarily distributed in the immune system. It is very important for the immune-modulatory effects of marijuana. In the long run, this study should also help us to develop better cannabinoid mimetics for the treatment of immune system illnesses, such as inflammation and autoimmune diseases. The drugs that soecifi(;allv taroeted at CB2 should be devoid of [}svchoactive side effects of marijuana.

这项研究计划的长期目标是确定以下功能的分子基础： CB2 大麻素受体。为实现这一目的，将采取以下步骤。在目标 1 和 2 中，将进行交互式诱变、分子建模和新化合物测试研究。这种组合方法增进了我们的理解，并且应该继续揭示关于大麻素配体如何被其受体识别，受体亚型选择性的起源，以及 _这些受体如何被激活/失活。大麻素受体共有许多保守的在其他经过充分研究的 G 蛋白偶联受体 (GPCR) 中发现的序列基序，例如视紫红质和生物胺受体。然而，几个螺旋弯曲残基和基序，以及一个关键的二硫键许多其他 GPCR 中存在的桥在大麻素受体中缺失。在目标 3 中，配体结合 CB2 大麻素受体的两个跨膜结构域的缝隙暴露将使用取代半胱氨酸可及性方法。这种方法应该阐明结构和功能大麻素受体的这些跨膜结构域的序列差异的后果。在目标4，CB2受体的二硫键形成将由最先进的技术直接确定质谱分析方法。研究独特的二硫键或二硫键的缺失键应提供CB2受体的重要螺旋折叠信息，这将具有重要意义关于配体结合缝隙和该受体激活机制的影响。最后，在目标 5、致力于大量功能性CB2受体的过表达和纯化。这具有挑战性的任务，如果成功完成，应该会通过提供糟糕的结果来产生重大影响需要纯受体蛋白用于未来的高分辨率生物物理研究。总的来说，这项研究应该帮助我们更详细地了解CB2受体的结构和功能。 CB2 是主要分布于免疫系统。对于免疫调节作用非常重要大麻。从长远来看，这项研究还应该有助于我们开发更好的大麻素模拟物治疗免疫系统疾病，例如炎症和自身免疫性疾病。那些药物 soecifi(;所有 CB2 的 taroeted 都应避免[}大麻的精神副作用。