Structure and Function of CB2 Cannabinoid Receptor

CB2 大麻素受体的结构和功能

• 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
• 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

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.

该研究建议的长期目标是确定分子基础的基础 CB2大麻素受体。为了实现此目的，将采取以下步骤。在AIM 1和2中 将进行互动诱变，分子建模和新的化合物测试研究。 这种结合的方法提高了我们的理解，应该继续揭示有关的新见解 如何通过其受体识别大麻素配体，受体亚型选择的起源和 这些受体如何被激活/灭活。大麻素受体共享许多保守的 在其他良好研究的G蛋白偶联受体（GPCR）中看到的序列基序，例如Rhodopsin和 生物胺受体。但是，几个螺旋弯曲的残基和图案以及一个关键的二硫化物 大麻素受体中缺少许多其他GPCR中的桥梁。在AIM 3中，配体绑定 CB2大麻素受体的两个跨膜结构域的缝隙暴露将使用 取代的半胱氨酸可及性法。这种方法应阐明结构和功能 大麻素受体的这些trasmemmbrane结构域中序列差异的后果。在 AIM 4，CB2受体的二硫键形成将直接通过最新出现确定 质谱法的方法论。研究独特的二硫键或二硫键 债券应提供CB2受体的重要螺旋折叠信息，这将具有重要的 该受体的配体结合缝隙和激活机制的含义。最后，目标 5，努力将用于过表达和净化大量功能CB2受体。这 具有挑战性的任务，如果成功完成，则应通过提供严重的影响产生重大影响 需要纯的受体蛋白，以进行未来的高分辨率生物物理研究。总体而言，这项研究应该 帮助我们以更分子的细节了解CB2受体的结构和功能。 CB2是 主要分布在免疫系统中。这对于免疫调节作用非常重要 大麻。从长远来看，这项研究还应帮助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。毒品 soecifi（;在CB2上打击的ALLV应没有[} svChoActive副作用的副作用。