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

基本信息

批准号：
7864275
负责人：
MARY E ABOOD
金额：
$ 31.77万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
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/7864275
关键词：
Adenylate Cyclase Affinity Agonist Amino Acids Binding CNR1 gene CNR2 gene Calcium Cannabinoids Cells Charge Communities Computer Simulation Computers Data Docking Drug Receptors Drug abuse Drug or chemical Tissue Distribution Endocannabinoids G-Protein-Coupled Receptors GTP-Binding Proteins Generations Goals 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 Models Molecular Profiling Mus Mutate Mutation Nature Neurons Organ Pathway interactions Peripheral Peripheral Nervous System Phosphotransferases Physiological Physiological Processes Property Proteins Pyrazoles Radiolabeled 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 design dioleoylphosphatidylcholine experience functional group human GPRC5C protein immunocytochemistry in vivo interest molecular dynamics molecular modeling mutant novel programs protein activation radiotracer receptor receptor binding receptor expression research study second messenger tool

项目摘要

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。这些作用大多数归因于大麻素CB1或CB2受体的作用。然而，显然没有CB1或CB2介导的效应可能不是受体介导的，但是也有足够的证据表明香草素受体（TRPV1）参与大麻素效应，至少另外两种其他大麻素受体亚型仅在目前为药理学定义。最近，已经建议两个孤儿G蛋白偶联受体（GPCR），GPR35和GPR55是大麻素受体，据报道，大麻素的配体据报道显示出每个大麻素受体。我们已经克隆并表达了GPR35和GPR55，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个GPCR。最近开发的GPR35和GPR55的计算机模型在其非活性和激活状态下将用于指导每个受体的突变研究。这些模型是通过我们对大麻素CB1和CB2受体的广泛建模和突变经验所告知的。突变研究的目的不仅是确定与配体识别有关的残基，而且对受体激活很重要的残基也是如此。在GPR35和GPR55配体识别研究中，将使用精心选择的化合物测试特定氨基酸与特定配体官能团的参与。体外突变研究的结果将用于完善计算机受体模型，以便在任何给定时间，这些模型反映了该领域的当前知识状态。因为迄今为止在GPR35和GPR55上的工作尚未确定任何受体的高亲和力拮抗剂，并且由于这些拮抗剂将是研究这两个受体的非常有价值的工具，因此拟议的工作的另一个目标是为每个受体亚型设计拮抗剂。这些化合物将在研究三角研究所（Seltzman）合成的UNCG（Reggio）设计，并在CPMCRI（Abood）进行了评估。从这项工作中出现的高亲和力拮抗剂将得到放射标记，并提供给科学界。确定GPR35和GPR35和GPR55的GPR35和信号转导途径的分布将有助于定义其生理和病理生理学作用。