New Tools for Studying Receptor Dimers

研究受体二聚体的新工具

• 批准号: 10579320
• 负责人: Paul J. Kammermeier
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579320
• 关键词: Affinity; Agonist; Amino Acid Sequence; Behavior; Biological; Brain; Brain region; Cell membrane; Cicatrix; Clinic; Cloning; Color; Communities; Complex; Data; Dimerization; Drug Screening; Endoplasmic Reticulum; Evaluation; Excision; Exhibits; Exteins; Family; Fluorescence Resonance Energy Transfer; Future; G-Protein-Coupled Receptors; Glutamates; Heterodimerization; Homo; Homodimerization; Knowledge; Libraries; Ligands; Link; Maps; Membrane; Membrane Proteins; Metabotropic Glutamate Receptors; Methods; Neurotransmitters; Oranges; Pathology; Performance; Pharmacology; Physiology; Population; Proteins; RNA Splicing; Receptor Signaling; Research; Shapes; Signal Transduction; Sirolimus; Site; Synaptic Transmission; System; Tail; Testing; Work; design; dimer; druggable target; glycoprotein 41; intein; metabotropic glutamate receptor 2; metabotropic glutamate receptor 4; notch protein; novel; pharmacologic; pre-clinical; receptor; receptor function; response; retention rate; sensor; targeted treatment; tool; trafficking

Abstract Metabotropic glutamate receptors (mGluRs) are class C G protein coupled receptors that function as dimers. While mGluRs are known to form homodimers, more recent work has shown that they can also heterodimerize, but not promiscuously. Because mGluRs exhibit widespread expression in the brain and regulate excitability and plasticity, they have become candidates as druggable targets for a variety of pathologies. To date however, excitement generated by preclinical data has not resulted in mGluR-targeting therapies in the clinic, despite a wealth of available ligands with good selectivity targeting these receptors. Our recent work examining mGluR2/4 heterodimers provides a possible explanation: ligands that are highly efficacious when targeting homodimeric receptors are often without effect when the same receptor is expressed as a heterodimer with another mGluR. Further complicating matters, these changes in pharmacological responses observed in mGluR2/4 heterodimers are not generalizable to all mGluR heterodimers, or even all mGluR2 containing heterodimers. Thus, to understand how any mGluR ligand will function in the brain, we must examine the pharmacological responses of each possible heterodimer pair in isolation. But this is complicated because every mGluR can also form homodimers, so any pair of expressed mGluR will have an unknown propensity to homo- and heterodimerize. To solve this problem, we have designed a novel dimer composition control system using a combination of ER retention sequences paired with orthogonal, split inteins, self-excising protein sequences, that will allow expression of pure populations of nearly wild type mGluR dimers of known composition. This system will be valuable not only in defining the pharmacological behavior of each dimer pair, but also by providing a broader and more accurate picture of the effects of mGluR targeting ligands by serving as a platform for broadly scoped drug screening. Further, this strategy provides a template for the study of other dimerizing membrane proteins. To this end, we will pursue the following Specific Aims: 1- Determine the performance and limitations of our novel eukaryotic split-intein system, and 2- Generate and test each mGluR combination with the split- intein system.

抽象的 代谢型谷氨酸受体（MGLURS）是C蛋白偶联受体，可作为二聚体。 尽管已知MGLUR会形成同二聚体，但最近的工作表明它们也可以异二聚体化，但 但不是滥交。因为mglurs在大脑中表现出广泛的表达并调节兴奋性和 可塑性，它们已成为候选人作为多种病理的可毒靶标。但是，到目前为止， 临床前数据产生的兴奋并未导致诊所中MGLUR靶向疗法，尽管 针对这些受体的大量可用配体具有良好的选择性。我们最近检查MGLUR2/4的工作 异二聚体提供了一种可能的解释：靶向同型二聚体时高效的配体 当相同的受体用另一种mglur表达同一受体时，受体通常无效。 进一步复杂的问题，在MGLUR2/4异二聚体中观察到的药理学反应的这些变化 不能推广到所有MGLUR异二聚体，甚至不可推广所有含有异二聚体的MGLUR2。因此， 了解任何mglur配体如何在大脑中起作用，我们必须检查药理反应 分离的每个可能的异二聚体对。但这很复杂，因为每个mglur也可以形成 同型二聚体，因此任何一对表达的MGlur都将具有同型和异二聚体的未知倾向。 为了解决这个问题，我们使用ER的组合设计了一种新型的二聚体组成控制系统 保留序列与正交的，分裂的蛋白，自抗蛋白质序列配对，这将允许 已知组成的几乎野生型mglur二聚体的纯种群的表达。这个系统将是 不仅可以定义每个二聚体对的药理学行为，而且通过提供更广泛的 更准确地描绘了MGLUR靶向配体的影响，通过充当广泛范围的平台 药物筛查。此外，该策略为研究其他二聚膜蛋白提供了模板。 为此，我们将追求以下具体目标：1-确定我们的绩效和局限性 新型真核生物分裂系统，2-生成和测试每个mglur的组合 内元系统。