Functional and Pharmacological Implications of mGluR Heteromerization

mGluR 异聚化的功能和药理学意义

• 批准号: 8452668
• 负责人: Paul J. Kammermeier
• 金额: $ 28.42万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-03 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8452668
• 关键词: Address; Affect; Agonist; American; Autistic Disorder; Binding; Biochemical; Biological Assay; Calcium Channel; Calcium Signaling; Cell fusion; Cells; Clinical; Color; Complementary DNA; Complex; Data; Dimerization; Disease; Drug Targeting; Electrophysiology (science); Epilepsy; Family; Fluorescence; Fluorescence Recovery After Photobleaching; G-Protein-Coupled Receptors; GTP-Binding Proteins; Glutamates; Hela Cells; Injection of therapeutic agent; Lead; Ligand Binding; Link; Literature; Mediating; Metabotropic Glutamate Receptors; Modeling; Molecular; Nervous system structure; Neuraxis; Neurons; Pain; Pathologic Processes; Pathology; Pathway interactions; Pharmacology; Phospholipase C; Physiological; Physiological Processes; Play; Property; Protein Family; Public Health; Rattus; Receptor Signaling; Role; Schizophrenia; Signal Transduction; Small Interfering RNA; Spectrum Analysis; Structure of superior cervical ganglion; System; Testing; Therapeutic; Transfection; base; dimer; innovation; interest; knock-down; member; multi-photon; novel; patch clamp; receptor; research study; stoichiometry; therapeutic target; tool

DESCRIPTION (provided by applicant): G protein coupled receptors (GPCRs) are the most frequently targeted protein family in clinical therapeutics. The molecular interactions between GPCRs can have a profound impact on their function and on the effects of drugs that target them. We examined the functional and pharmacological effects of interactions between two glutamate-activated GPCRs called mGluR1 and mGluR5, which are known to form homodimeric complexes (mGluR1-mGluR1 and mGluR5-mGluR5 dimers), are involved in many patho-physiological processes, and have widespread distribution in the central nervous system. Our data suggest that these receptors can interact functionally, and that this interaction alters their sensitivity to selective pharmacological agents that target them. We suggest a novel hypothesis to explain this interaction in which mGluR1 and mGluR5 dimers combine to form tetrameric (or possibly larger) complexes with the ability to trans-activate, such that ligand binding to one receptor may activate G protein through the other. Further, these putative tetrameric receptors seem to respond to some pharmacological agents differently than their homodimeric constituents. To address this hypothesis, we will pursue the following Specific Aims: 1) To characterize the unique pharmacological properties of mGluR1/5 heteromers using the rat superior cervical ganglion (SCG) neuronal expression system, 2) To confirm the pharmacological and signaling properties of mGluR1/5 heteromers by expression in HeLa cells and using a different effectors to assay receptor signaling, 3) To directly assess the ability of fluorescently-tagged mGluR1, mGluR5 and both receptors together to form dimeric, tetrameric, or higher order multimeric receptors using multi-photon Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS), and 4) To verify the physiological relevance of functional mGluR interactions in neurons that natively express both mGluR1 and mGluR5.

描述（由申请人提供）：G蛋白偶联受体（GPCR）是临床疗法中最常见的靶向蛋白质家族。 GPCR之间的分子相互作用可以对它们的功能以及针对它们的药物的影响产生深远的影响。我们研究了称为Mglur1和Mglur5的两个谷氨酸激活的GPCR之间相互作用的功能和药理效应，已知它们形成同型二聚体复合物（MGLUR1-MGLUR1和MGLUR5-MGLUR5二聚体），与许多病理学过程中的分布相关，并且具有广泛的分布。我们的数据表明这些受体可以在功能上相互作用，并且这种相互作用改变了它们 对目标的选择性药理学剂的敏感性。我们提出了一种新的假设来解释这种相互作用，其中mGlUR1和MGLUR5二聚体结合形成四聚体（或可能更大的）复合物具有反式激活的能力，因此配体与一种受体的结合可以通过另一个受体激活G蛋白。此外，这些推定的四聚体受体似乎对某些药理剂的反应与同二聚体成分不同。为了解决这一假设，我们将追求以下具体目的：1）使用大鼠上颈神经节（SCG）神经元表达系统来表征MGLUR1/5异构体的独特药理特性，2）确认以外的效率效应的MGLUR1/5杂体的药理和信号的效率，以确认药物学和信号的特性，以使效率效应效率效应，以实现3个效率的效率，以实现3个效率的效率，并使用效率效应效应效应。 fluorescently-tagged mGluR1, mGluR5 and both receptors together to form dimeric, tetrameric, or higher order multimeric receptors using multi-photon Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS), and 4) To verify the physiological relevance of functional mGluR interactions in neurons that natively express both mGluR1 and mglur5。