Molecular Mechanisms of Synaptic G Protein-Coupled Receptors

突触G蛋白偶联受体的分子机制

• 批准号: 10166865
• 负责人: Joshua Levitz
• 金额: $ 41.91万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-07-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10166865
• 关键词: Area; Biophysics; Coupling; Disease; Drug Targeting; Environment; Fluorescence Resonance Energy Transfer; G-Protein-Coupled Receptors; GABA Receptor; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Glutamates; Goals; Health; Hippocampus (Brain); Kinetics; Lead; Ligand Binding; Ligand Binding Domain; Ligands; Link; Long-Term Depression; Measurement; Measures; Membrane; Mental disorders; Metabotropic Glutamate Receptors; Methods; Molecular; Molecular Conformation; Motion; Nervous system structure; Neurotransmitters; Optical Methods; Optics; Pattern; Pharmacology; Population; Process; Receptor Activation; Reporter; Research; Rhodopsin; Role; Signal Transduction; Specificity; Structure; Synapses; Synaptic Transmission; Transmembrane Domain; Work; beta-adrenergic receptor; biological systems; desensitization; detector; dimer; extracellular; glutamatergic signaling; improved; insight; nervous system disorder; neuronal excitability; neurotransmitter release; optical sensor; optogenetics; protein activation; receptor; response; single molecule; spatiotemporal

Project Summary In many biological systems G protein-coupled receptors (GPCRs) provide a crucial molecular link between the dynamics of the extracellular environment and the associated intracellular signaling response. In the nervous system, GPCRs serve as detectors of precise patterns of neurotransmitter release and are able to, in turn, modulate neuronal excitability and synaptic transmission. Of particular importance are the class C metabotropic glutamate (mGluR) and GABA receptors (GABABR), which respond to the major excitatory and inhibitory neurotransmitters, respectively, and serve as drug targets for neurological and psychiatric disorders. Unfortunately, our understanding of their underlying molecular mechanisms of signaling remain limited due to a lack of methods for the direct measurement and manipulation of their activity with high specificity and spatial and temporal precision. Furthermore, the biophysical activation mechanism of class C GPCRs is particularly challenging to decipher because unlike class A GPCRs, such as rhodopsin or ß-adrenergic receptors, they contain large, extracellular ligand binding domains (LBDs) that multimerize and couple, via a poorly understood mechanism, to a transmembrane domain (TMD). Our recent work has established new optical methods for directly measuring mGluR assembly and conformational dynamics at the single molecule level and has also produced an optogenetic method to manipulate receptors with subtype selectivity and high spatiotemporal precision using photoswitchable tethered ligands. These breakthroughs have advanced our understanding of how mGluRs dimerize and the initial molecular motions that lead to cooperative receptor activation, but many fundamental questions remain. In research area 1 we will dissect the activation mechanism of mGluRs and GABABRs in a quantitative, interdisciplinary way using optical approaches, including single molecule Forster resonance energy transfer (FRET) to measure conformational dynamics, in conjunction with functional reporters and detailed structural analysis. The long-term goal is to understand, biophysically, how allosteric inter-domain and inter-subunit coupling interactions permit orthosteric and allosteric ligand binding to produce G protein activation. This work will give major insight into the fundamental activation processes of a large class of membrane receptors and should provide a deeper understanding of their molecular pharmacology. In research area 2 we will improve and harness the power of optical sensors of activation and optogenetic control of receptors to probe the kinetics of different mGluR subtypes at the level of activation, signaling, and desensitization and to dissect their spatiotemporal signaling profiles at hippocampal synapses. In the long term we plan to use this information to probe the mechanism of induction of long-term depression by pre-synaptic, post-synaptic, and glial mGluR populations. This work will provide a dynamic picture of mGluR signaling that has been missing from the field and will strengthen our molecular understanding of the role of these receptors in synaptic modulation in health and disease.

项目总结

项目成果

Optogenetic Techniques for Manipulating and Sensing G Protein-Coupled Receptor Signaling.

• DOI: 10.1007/978-1-0716-0755-8_2
• 发表时间: 2020
• 期刊: Methods in molecular biology (Clifton, N.J.)

Distinct beta-arrestin coupling and intracellular trafficking of metabotropic glutamate receptor homo- and heterodimers.

• DOI: 10.1126/sciadv.adi8076
• 发表时间: 2023-12-08
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Lee, Joon;Gonzalez-Hernandez, Alberto J.;Kristt, Melanie;Abreu, Nohely;Rossmann, Kilian;Arefin, Anisul;Marx, Dagan C.;Broichhagen, Johannes;Levitz, Joshua
• 通讯作者: Levitz, Joshua

Glutamatergic Signaling in the Central Nervous System: Ionotropic and Metabotropic Receptors in Concert.

中枢神经系统中的谷氨酸能信号传导：共同的离子和代谢型受体。

• DOI: 10.1016/j.neuron.2018.05.018
• 发表时间: 2018-06-27
• 期刊: Neuron
• 影响因子: 16.2
• 作者: Reiner A;Levitz J
• 通讯作者: Levitz J

Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin.

• DOI: 10.1038/s41598-021-90039-3
• 发表时间: 2021-05-18
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Khelashvili G;Pillai AN;Lee J;Pandey K;Payne AM;Siegel Z;Cuendet MA;Lewis TR;Arshavsky VY;Broichhagen J;Levitz J;Menon AK
• 通讯作者: Menon AK

Interrogating surface versus intracellular transmembrane receptor populations using cell-impermeable SNAP-tag substrates.

• DOI: 10.1039/d0sc02794d
• 发表时间: 2020-07-07
• 期刊: Chemical science
• 影响因子: 8.4
• 作者: Poc P;Gutzeit VA;Ast J;Lee J;Jones BJ;D'Este E;Mathes B;Lehmann M;Hodson DJ;Levitz J;Broichhagen J
• 通讯作者: Broichhagen J