Molecular Characterization of GPR35 and GPR55, Putative Cannabinoid Receptors

GPR35 和 GPR55（假定的大麻素受体）的分子表征

• 批准号: 7646619
• 负责人: MARY E ABOOD
• 金额: $ 34.41万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7646619
• 关键词: Adenylate Cyclase; Affinity; Agonist; Amino Acids; Binding; CNR1 gene; CNR2 gene; Calcium; Cannabinoids; Cells; Charge; Class; Communities; Computer Simulation; Computers; Condition; Data; Docking; Drug Receptors; Drug abuse; Drug or chemical Tissue Distribution; Endocannabinoids; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Goals; Helix (Snails); Herb; In Vitro; Indoles; Institutes; Knowledge; Ligand Binding; Ligands; Literature; Marijuana; Mediating; Memory; Messenger RNA; Microglia; Mitogen-Activated Protein Kinases; Modeling; Molecular; Molecular Conformation; Molecular Profiling; Mus; Mutate; Mutation; Nature; Neurons; Numbers; Organ; Pathway interactions; Peripheral; Peripheral Nervous System; Phosphotransferases; Physiological; Physiological Processes; Property; Proteins; Pyrazoles; Radiolabeled; Range; Receptor Activation; Regulation; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; SR 141716A; Scheme; Second Messenger Systems; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Simulate; Site; Site-Directed Mutagenesis; Structure; TRPV1 gene; Techniques; Testing; Time; Transmembrane Domain; Western Blotting; Work; base; cannabinoid receptor; capsaicin receptor; computer generated; design; dioleoylphosphatidylcholine; drug mechanism; ear helix; experience; functional group; human GPRC5C protein; immunocytochemistry; in vivo; indole; interest; molecular dynamics; molecular modeling; mutant; novel; programs; protein activation; pyrazole; radiotracer; receptor; receptor binding; receptor expression; research study; second messenger; tool

DESCRIPTION (provided by applicant): A large number of physiological processes are controlled by the endogenous cannabinoids. Most of these effects have been attributed to action at either the cannabinoid CB1 or CB2 receptors. Yet there are effects that clearly are not CB1- or CB2-mediated which may not be receptor mediated, but there is also sufficient evidence to suggest the involvement of the vanilloid receptor (TRPV1) in cannabinoid effects and at least two other cannabinoid receptor subtypes defined only pharmacologically until now. Very recently, two orphan G-protein coupled receptors (GPCRs), GPR35 and GPR55, have been suggested to be cannabinoid receptors, with a fairly wide range of cannabinoid ligands reported to display affinity/efficacy at each. We have cloned and expressed both GPR35 and GPR55 and our preliminary studies confirm that they are activated by multiple cannabinoid compounds. In work proposed here, we plan to characterize these two GPCRs through ligand binding and functional studies. Recently developed computer models of GPR35 and GPR55 in their inactive and activated states will be used to guide mutation studies of each receptor. These models are informed by our extensive modeling and mutation experience with the cannabinoid CB1 and CB2 receptors. The goal of mutation studies will be not only to identify residues involved in ligand recognition, but also those residues important for receptor activation. In GPR35 and GPR55 ligand recognition studies, the involvement of specific amino acids with specific ligand functional groups will be tested using carefully chosen compounds. Results of mutation studies in vitro will be used to refine computer receptor models, such that at any given time, these models reflect the current state of knowledge in the field. Because work thus far on GPR35 and GPR55 has not identified a high affinity antagonist for either receptor and because such antagonists would be very valuable tools for studying these two receptors, another goal of the proposed work will be to design antagonists for each receptor sub-type. These compounds will be designed at UNCG (Reggio), synthesized at Research Triangle Institute (Seltzman), and evaluated at CPMCRI (Abood). High affinity antagonists that emerge from this work will be radiolabeled and made available to the scientific community. Determining the distribution of GPR35 and signal transduction pathways of GPR35 and GPR55 will help define their physiological and pathophysiological roles.

描述（由申请人提供）：大量生理过程由内源性大麻素控制。这些效应中的大多数归因于大麻素CB 1或CB 2受体的作用。然而，也有明显不是CB 1或CB 2介导的效应，可能不是受体介导的，但也有足够的证据表明香草素受体（TRPV 1）参与大麻素效应，至少还有两种其他大麻素受体亚型迄今为止仅被定义。最近，两种孤儿G蛋白偶联受体（GPCR），GPR 35和GPR 55，已被认为是大麻素受体，有相当广泛的大麻素配体报告显示在每个亲和力/功效。我们已经克隆并表达了GPR 35和GPR 55，我们的初步研究证实它们被多种大麻素化合物激活。在这里提出的工作中，我们计划通过配体结合和功能研究来表征这两种GPCR。最近开发的计算机模型GPR 35和GPR 55在其非活性和激活状态将用于指导每个受体的突变研究。这些模型是由我们对大麻素CB 1和CB 2受体的广泛建模和突变经验提供的。突变研究的目标不仅是识别参与配体识别的残基，而且是识别对受体活化重要的残基。在GPR 35和GPR 55配体识别研究中，将使用精心选择的化合物检测特定氨基酸与特定配体官能团的关系。体外突变研究的结果将用于改进计算机受体模型，以便在任何给定时间，这些模型反映该领域的当前知识状态。由于迄今为止对GPR 35和GPR 55的工作尚未鉴定出对任一受体的高亲和力拮抗剂，并且由于这种拮抗剂将是研究这两种受体的非常有价值的工具，因此所提出的工作的另一个目标将是设计每种受体亚型的拮抗剂。这些化合物将在UNCG（Reggio）设计，在Research Triangle Institute（Seltzman）合成，并在CPMCRI（Pennsylvania）进行评估。从这项工作中产生的高亲和力拮抗剂将被放射性标记，并提供给科学界。确定GPR 35的分布以及GPR 35和GPR 55的信号转导途径将有助于确定其生理和病理生理作用。