Structural insight into TRPA1 channel interactionwith agonist and antagonist

TRPA1 通道与激动剂和拮抗剂相互作用的结构洞察

• 批准号: 9225310
• 负责人: Vera Moiseenkova-Bell
• 金额: $ 1.91万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9225310
• 关键词: Agonist; Analgesics; Anti-Inflammatory Agents; Architecture; Attention; Binding; Cations; Chemicals; Cryoelectron Microscopy; Cysteine; Data; Disulfides; Docking; Electron Microscopy; Electrophysiology (science); Elements; Esthesia; Grant; Health; Homology Modeling; Inflammation Mediators; Ion Channel; Knowledge; Laboratories; Length; Ligands; Mammals; Mass Spectrum Analysis; Mediating; Methods; Modification; Molecular; Molecular Conformation; Molecular Models; N-terminal; Oximes; Pain; Pain management; Proteolysis; Refractory; Resolution; Rest; Role; Sensory; Signal Transduction; Site; Site-Directed Mutagenesis; Structure; Sulfhydryl Compounds; Tertiary Protein Structure; Testing; Therapeutic; Transmembrane Domain; Work; base; density; desensitization; design; disulfide bond; emotional experience; flexibility; in vivo; inflammatory neuropathic pain; inflammatory pain; inhibitor/antagonist; insight; molecular modeling; novel; pre-clinical trial; receptor; response; sensor

DESCRIPTION (provided by applicant): TRPA1 is a Ca2+-permeable, non-selective cation channel and one of the key pain sensors in mammals. It has emerged as a novel target for analgesics and anti-inflammatory agents. Pain sensation mediated by TRPA1 involves modification of N-terminal cysteine residues on the channel by thiol-reactive compounds and inflammatory mediators. Binding of thiol-reactive compounds to the channel in the resting state leads to channel activation followed rapidly by desensitization. While channel activity has been studied extensively by electrophysiological methods, little is known about the structural mechanisms of channel activation and desensitization. Additionally, two compounds that directly inhibit TRPA1 are in pre-clinical trials, and understanding the mechanism of TRPA1 inhibition by these compounds will be important for advancing our knowledge of the structural differences between functionally relevant TRPA1 conformations. Using cryo-EM, site-directed mutagenesis, limited proteolysis, and mass spectrometry, in this grant we aim to establish a detailed structural understanding of the mechanisms of TRPA1 channel activation, desensitization and inhibition, which will help facilitate rational design of novel analgesics.

描述（由申请人提供）：TRPA1是Ca2+ - 可渗透的，非选择性的阳离子通道，也是哺乳动物中的关键疼痛传感器之一。它已成为镇痛药和抗炎药的新靶标。 TRPA1介导的疼痛感涉及通过硫醇反应性化合物和炎症介质对通道上N末端半胱氨酸残基的改性。硫醇反应化合物与静息状态中通道的结合导致通道激活，然后迅速脱敏。尽管通过电生理方法对通道活性进行了广泛的研究，但对通道激活和脱敏的结构机制知之甚少。此外，在临床前试验中，两种直接抑制TRPA1的化合物是在临床前试验中，而这些化合物抑制TRPA1的机制对于促进我们对功能相关的TRPA1构象之间的结构差异的了解至关重要。使用冷冻EM，定向诱变，有限的蛋白水解和质谱法，我们旨在建立详细的结构 了解TRPA1通道激活，脱敏和抑制的机制，这将有助于促进新型镇痛药的理性设计。