Alpha7 Nicotinic Receptor: Structures and Coupling with Intracellular Proteins

Alpha7 烟碱受体：结构及其与细胞内蛋白质的偶联

• 批准号: 10393547
• 负责人: PEI TANG
• 金额: $ 47.88万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10393547
• 关键词: Action Potentials; Address; Affect; Agonist; Binding; Binding Sites; Biophysics; Brain; Cell surface; Chimera organism; Cognition; Cognitive; Coupling; Cryoelectron Microscopy; Crystallization; Development; Disease; Down-Regulation; Drug Targeting; Electron Spin Resonance Spectroscopy; Extracellular Domain; Extracellular Structure; Foundations; Graph; Human; Immune system; Investigation; Ion Channel Gating; Learning; Left; Length; Ligands; Mediating; Memory; Modeling; Molecular; Molecular Conformation; NMR Spectroscopy; Neurodegenerative Disorders; Neurons; Nicotinic Receptors; Outcomes Research; Play; Proteins; Regulation; Resolution; Rest; Roentgen Rays; Role; Scaffolding Protein; Side; Signal Pathway; Signal Transduction; Site; Solid; Structure; Subcellular structure; Synapses; Synaptic plasticity; Therapeutic; Transmembrane Domain; X-Ray Crystallography; alpha-bungarotoxin receptor; base; desensitization; design; experimental study; extracellular; flexibility; nervous system disorder; neurotransmission; novel; positive allosteric modulator; rational design; receptor; restraint; targeted treatment; therapeutic target; trafficking; transmission process; treatment strategy

Neuronal nicotinic acetylcholine receptors (nAChRs) belong to the superfamily of pentameric ligand-gated ion channels (pLGICs) that mediate fast neurotransmission and are therapeutic targets for various neurological diseases and disorders. However, no single high-resolution structure has yet been determined for any full-length nAChRs that contains the extracellular (ECD), transmembrane (TMD), and intracellular (ICD) domains. Neither X-ray crystallography nor cryo-EM is suitable for resolving structures of the flexible ICD. Lines of evidence suggest direct interactions between nAChRs and intracellular proteins implicated in synaptic plasticity and cell signaling pathways, but few molecular details have been revealed due to the lack of high-resolution structures of the ICD. Here, we propose to determine structures of α7nAChR, which is one of the most abundant nAChR subtypes in the brain. We have successfully produced functional human full-length α7nAChR and the TMD+ICD of α7nAChR along with substantial preliminary structural and functional results, which built a solid foundation for carrying out structural investigations of α7nAChR. Using nuclear magnetic resonance spectroscopy (NMR) and electron paramagnetic resonance (EPR), we will determine atomic-resolution structures of the ICD. The ICD structures will be integrated with available structures of the ECD and TMD, as well as with our newly collected structure restraints to generate structures for full-length α7nAChR in the resting state. We will elucidate conformational changes underlying α7nAChR desensitization, which is known to play a significant role in the synaptic control of action-potential transmission. Finally, we will determine binding modes of α7nAChR networked with intracellular scaffold proteins that affect the distribution of α7nAChR on the cell surface. The generated structures will provide useful information for developing treatment strategies for diseases related to α7nAChR.

神经元烟碱型乙酰胆碱受体（nAChRs）属于五聚体配体门控离子受体超家族 通道（pLGIC）介导快速神经传递，是各种神经系统疾病的治疗靶点。 疾病和失调。然而，还没有确定任何单一的高分辨率结构， 全长nAChR，包含细胞外（ECD）、跨膜（TMD）和细胞内（ICD） 域. X射线晶体学和冷冻EM都不适合解析柔性ICD的结构。线 的证据表明nAChRs和参与突触的细胞内蛋白质之间的直接相互作用， 可塑性和细胞信号传导途径，但由于缺乏 ICD的高分辨率结构。在这里，我们建议确定α7nAChR的结构，这是一个 最丰富的nAChR亚型。我们已经成功生产出功能正常的人体全长 α 7 nAChR和α 7 nAChR的TMD+ICD沿着，具有实质性的初步结构和功能结果， 为进一步研究α7nAChR的结构奠定了基础。使用核磁 共振光谱（NMR）和电子顺磁共振（EPR），我们将确定 ICD的原子分辨率结构。疾病分类结构将与 ECD和TMD，以及我们新收集的结构约束，以生成全长结构 静息状态下的α 7 nAChR。我们将阐明α 7 nAChR脱敏的基础构象变化， 已知其在动作电位传递的突触控制中起重要作用。最后我们将 确定α 7 nAChR与细胞内支架蛋白联网的结合模式，这些蛋白影响细胞内 细胞表面的α 7 nAChR。所生成的结构将为开发治疗方法提供有用的信息 与α7nAChR相关的疾病的策略。

项目成果

Structural Elucidation of Ivermectin Binding to α7nAChR and the Induced Channel Desensitization.

• DOI: 10.1021/acschemneuro.2c00783
• 发表时间: 2023-03-15
• 期刊: ACS CHEMICAL NEUROSCIENCE
• 影响因子: 5
• 作者: Bondarenko, Vasyl;Chen, Qiang;Singewald, Kevin;Haloi, Nandan;Tillman, Tommy S.;Howard, Rebecca J.;Lindahl, Erik;Xu, Yan;Tang, Pei
• 通讯作者: Tang, Pei

SARS-CoV-2 Spike Protein Downregulates Cell Surface α7nAChR through a Helical Motif in the Spike Neck.

• DOI: 10.1021/acschemneuro.2c00610
• 发表时间: 2023-02-15
• 期刊: ACS CHEMICAL NEUROSCIENCE
• 影响因子: 5
• 作者: Tillman, Tommy S.;Chen, Qiang;Bondarenko, Vasyl;Coleman, Jonathan A.;Xu, Yan;Tang, Pei
• 通讯作者: Tang, Pei