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

基本信息

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
项目类别：
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

项目摘要

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.

神经元烟碱乙酰胆碱受体（NACHRS）属于五聚体门控离子的超家族介导快速神经传递的渠道（PLGIC）是各种神经系统的治疗靶标疾病和疾病。但是，尚未确定任何高分辨率结构包含细胞外（ECD），跨膜（TMD）和细胞内（ICD）的全长NACHRS 域。 X射线晶体学和冷冻EM都不适合柔性ICD的结构。线证据表明NACHRS与突触中实现的细胞内蛋白之间的直接相互作用可塑性和细胞信号通路，但由于缺乏分子细节，很少有分子细节 ICD的高分辨率结构。在这里，我们建议确定α7NACHR的结构，这是其中之一大脑中最丰富的NACHR亚型。我们已经成功产生了功能性的人类全长 α7NACHR和α7NACHR的TMD+ICD以及实质性的初步结构和功能结果，这为进行α7NACHR的结构研究奠定了坚实的基础。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。 ICD结构将与可用结构集成 ECD和TMD，以及我们新收集的结构约束，以生成完整长度的结构 α7nachr处于静止状态。我们将阐明α7nAChR脱敏的基础构象变化，众所周知，这在动作电位传播的突触控制中起着重要作用。最后，我们会的确定与细胞内支架蛋白网络网络的α7NACHR的结合模式，这些模式影响影响的分布细胞表面上的α7NACHR。生成的结构将为开发治疗提供有用的信息与α7NACHR有关的疾病策略。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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