Spectroscopic analyses of TRPV1 during gating

门控过程中 TRPV1 的光谱分析

• 批准号: 10039442
• 负责人: Julio F Cordero-Morales
• 金额: $ 43.22万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10039442
• 关键词: Afferent Neurons; Aging; Analgesics; Animals; Arthritis; Binding Sites; Bradykinin; C-terminal; Capsaicin; Cartoons; Cell membrane; Charge; Chemicals; Chili Pepper; Code; Cryoelectron Microscopy; Cysteine; Data; Detection; Development; Drug Design; Electron Microscopy; Electron Spin Resonance Spectroscopy; Electrophysiology (science); Goals; Hydrolysis; Individual; Inflammation; Inflammatory; Ion Channel; Light; Link; Lipids; Liposomes; Malignant Neoplasms; Measures; Mediating; Membrane; Membrane Lipids; Membrane Proteins; Mission; Molecular; Molecular Conformation; Nervous system structure; Neurogenic Inflammation; Neurons; Outcome; Pain; Pain management; Pathway interactions; Pharmacology; Phosphatidylinositol 4,5-Diphosphate; Phospholipase; Play; Process; Protons; Public Health; Rattus; Research; Role; Secondary to; Sensory Physiology; Site; Solvents; Stimulus; Structure; Syndrome; TRP channel; TRPV channel; Techniques; Testing; Toxin; United States National Institutes of Health; Vanilloid; base; design; experimental study; extracellular; inflammatory pain; mutant; novel; particle; protonation; receptor; reconstitution; response; stimulus sensitivity; tissue injury; tool

The transient receptor potential vanilloid 1 (TRPV1) is a polymodal ion channel essential to the cellular mechanism underlying the detection of noxious stimuli. TRPV1 is activated by heat, protons, capsaicin, and animal toxins, and is modulated by proalgesic inflammatory agents (e.g., bradykinin, bioactive lipids) produced in response to tissue injury. Our long-term goal is to delineate the roles of polymodal ion channels in sensory neuron excitation and the mechanisms by which they contribute to inflammatory pain. The rationale for our proposed research is that a deeper mechanistic understanding of TRPV1 proton- and heat-dependent gating would greatly facilitate the development of strategies to ameliorate TRPV1-mediated inflammatory pain, without disrupting normal sensory physiology. While functional and structural characterization of TRPV1 have shed light on the mechanisms of capsaicin and toxin activation, the processes whereby the two main endogenous activators, protons and heat, trigger gating remain largely unknown. Moreover, the intracellular TRPV1 C terminus is a key regulatory site for regulating stimulus sensitivity. However, any potential allosteric interacting regions or putative contacts with the plasma membrane have yet remain to be explored. It is our contention that spectroscopic approaches are needed to fully define the allosteric conformational changes responsible for TRPV1 activation and to depict the C-terminal/membrane interaction. To this end, we will carry out electrophysiological analyses together with electron paramagnetic resonance spectroscopy experiments in both closed and open states. With these data, we will depict the conformational changes that TRPV1 undergoes during proton- and heat-dependent gating. We will pursue two Specific Aims: 1) Determine the dynamic conformational rearrangements of TRPV1 during proton and heat activation, and 2) Explore the interaction between the TRPV1 C-terminal domain and the plasma membrane. The proposed research is significant because it is expected to have broad translational importance in the treatment of pain associated with a wide range of pathophysiological conditions.

瞬时受体电位香草素1(TRPV1)是一种细胞内必需的多峰离子通道。 检测有害刺激的潜在机制。TRPV1被热、质子、辣椒素和 动物毒素，由产生的致痛性炎症剂(如缓激肽、生物活性脂)调节 以应对组织损伤。我们的长期目标是描述多模式离子通道在感觉中的作用 神经元兴奋及其导致炎症性疼痛的机制。我们的理由是 提出的研究是对TRPV1质子和热依赖门控的更深层次的机理理解 将极大地促进制定缓解TRPV1介导的炎性疼痛的策略，而不需要 扰乱了正常的感觉生理学。虽然TRPV1的功能和结构特征已经阐明 辣椒素和毒素激活的机制，两种主要内源物质激活的过程 激活剂，质子和热，触发门控在很大程度上仍然是未知的。此外，细胞内的TRPV1C 终末是调节刺激敏感性的关键调控部位。然而，任何潜在的变构相互作用 与质膜的区域或可能的接触仍有待探索。我们的论点是 需要光谱学方法来充分定义引起变构构象变化 TRPV1的激活和C末端/膜的相互作用。为此，我们将开展 两者的电生理分析和电子顺磁共振波谱实验 封闭和开放状态。有了这些数据，我们将描述TRPV1经历的构象变化 在依赖质子和热量的选通过程中。我们将追求两个具体目标：1)确定动态 TRPV1在质子和热激活过程中的构象重排，以及2)相互作用的探讨 在TRPV1C-末端结构域和质膜之间。这项拟议的研究具有重要意义 因为预计它在治疗与广泛相关的疼痛方面具有广泛的翻译重要性 病理生理条件的范围。

项目成果

Protein functional dynamics from the rigorous global analysis of DEER data: Conditions, components, and conformations.

• DOI: 10.1085/jgp.201711954
• 发表时间: 2021-11-01
• 期刊: The Journal of general physiology
• 作者: Hustedt EJ;Stein RA;Mchaourab HS
• 通讯作者: Mchaourab HS

Deficiency of Inositol Monophosphatase Activity Decreases Phosphoinositide Lipids and Enhances TRPV1 Function In Vivo.

• DOI: 10.1523/jneurosci.0803-20.2020
• 发表时间: 2021-01-20
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Caires R;Bell B;Lee J;Romero LO;Vásquez V;Cordero-Morales JF
• 通讯作者: Cordero-Morales JF