Spectroscopic analyses of TRPV1 during gating

门控过程中 TRPV1 的光谱分析

基本信息

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
项目类别：
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

项目摘要

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.

瞬时受体电位香草酸 1 (TRPV1) 是细胞必需的多模式离子通道检测有害刺激的机制。 TRPV1 被热、质子、辣椒素和动物毒素，并由产生的促痛炎症剂（例如缓激肽、生物活性脂质）调节以应对组织损伤。我们的长期目标是描绘多模式离子通道在感官中的作用神经元兴奋及其导致炎性疼痛的机制。我们的理由拟议的研究是对 TRPV1 质子和热依赖性门控的更深入的机制理解将极大地促进改善 TRPV1 介导的炎症疼痛策略的开发，而无需扰乱正常的感觉生理机能。 TRPV1 的功能和结构表征已经揭示了关于辣椒素和毒素激活的机制，两种主要内源性的过程激活剂、质子和热，触发门控仍然很大程度上未知。此外，细胞内 TRPV1 C 末端是调节刺激敏感性的关键调节位点。然而，任何潜在的变构相互作用区域或与质膜的假定接触仍有待探索。我们的论点是需要光谱方法来充分定义导致的变构构象变化 TRPV1 激活并描述 C 端/膜相互作用。为此，我们将开展电生理分析以及电子顺磁共振波谱实验关闭和开放状态。利用这些数据，我们将描述 TRPV1 经历的构象变化在质子和热依赖性门控期间。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) 探索相互作用 TRPV1 C 末端结构域和质膜之间。拟议的研究意义重大因为预计它在治疗与广泛相关的疼痛方面具有广泛的转化重要性病理生理状况的范围。

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）

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

肌醇单磷酸酶活性缺乏会降低体内磷酸肌醇脂质并增强 TRPV1 功能。

DOI：
发表时间：
2021
期刊：
影响因子：
0
作者：
Caires, Rebeca;Bell, Briar;Lee, Jungsoo;Romero, Luis O;Vásquez, Valeria;Cordero
通讯作者：
Cordero

A dietary fatty acid counteracts neuronal mechanical sensitization.

膳食脂肪酸可以抵消神经元机械敏化。

DOI：
发表时间：
2020-06-19
期刊：
影响因子：
16.6
作者：
Romero, Luis O;Caires, Rebeca;Nickolls, Alec R;Chesler, Alexander T;Cordero;Vásquez, Valeria
通讯作者：
Vásquez, Valeria

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

来自对 DEER 数据的严格全局分析的蛋白质功能动力学：条件、成分和构象。