Mechanisms of TRPV1 channel gating and modulation by temperature and vanilloids

TRPV1 通道门控和温度和香草酸调节机制

• 批准号: 10467443
• 负责人: Andres Jara Oseguera
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10467443
• 关键词: Mechanisms; TRPV1; channel; gating; modulation

My long-term goal is to establish an independent research group aimed at understanding allosteric signaling in membrane proteins. I intend to focus on Transient Receptor Potential (TRP) cation channels as a model system, as they exhibit fascinating allosteric behaviors that have important consequences for diverse physiological processes. One of the best characterized TRP channels is the TRPV1. The activation of this channel in nociceptor terminals by noxious environmental stimuli, including burning heat, leads directly to the perception of pain, and can contribute to the development of inflammatory hyperalgesia. All these processes ultimately depend on the opening and closing (i.e. gating) of the cation-conduction pathway in the pore of TRPV1, and its control by positive and negative modulators. We currently lack a mechanistic understanding of the process of TRP channel gating, as well as of the mechanisms through which gating is controlled by important stimuli such as temperature and vanilloids. This project will first aim to identify the regions of the channel that directly control gating of the pore. Second, it will aim at determining whether the extracellular half of the pore forms the temperature sensor. Third, it will aim at elucidating the mechanisms through which distinct vanilloid compounds can either activate or inhibit the TRPV1 channel. This has important consequences for the effects of TRPV1-targetted analgesic drugs on thermoregulation, and for the function of endogenous pro-inflammatory activators of TRPV1. Finally, I will engineer sensitivity to vanilloids into the TRPV2 channel and study whether the mechanisms of channel modulation by these molecules are conserved between V1 and V2. The three aims will be addressed with a combination of electrophysiological measurements of channel activity, site-directed mutagenesis, computational biophysics and structural information. The mentored part of the award will allow me to obtain the necessary training to understand computational biophysics approaches and rigorously apply them to my future research. This will complement my training as an experimental physiologist, providing me with an extended conceptual framework and a set of practical tools to investigate protein function at the level of structural dynamics at atomic scales. I consider that the integration of protein structure, function and dynamics is necessary to fully comprehend the mechanisms that govern allosterism in proteins, and I therefore consider that the training that I will obtain will be fundamental for my career. In addition, training in the Swartz lab, one of the leading laboratories worldwide in ion channel research, will strengthen my expertise in electrophysiology and broaden my overall scientific perspective.

我的长期目标是建立一个独立的研究小组，旨在了解膜蛋白中的变构信号传导。我打算重点关注瞬时受体电位 (TRP) 阳离子通道作为模型系统，因为它们表现出令人着迷的变构行为，对不同的生理过程产生重要影响。 TRPV1 是最具特征的 TRP 通道之一。伤害性环境刺激（包括灼热）激活伤害感受器末梢的该通道，直接导致疼痛感，并可能导致炎症性痛觉过敏的发生。所有这些过程最终取决于TRPV1孔内阳离子传导途径的打开和关闭（即门控），以及正负调节剂的控制。目前，我们对 TRP 通道门控过程以及温度和香草醛等重要刺激控制门控的机制缺乏了解。该项目首先旨在确定直接控制孔门的通道区域。其次，其目的是确定孔的细胞外部分是否形成温度传感器。第三，它将旨在阐明不同的香草酸化合物激活或抑制 TRPV1 通道的机制。这对于 TRPV1 靶向镇痛药物对体温调节的影响以及 TRPV1 内源性促炎激活剂的功能具有重要影响。最后，我将对香草类化合物的敏感性设计到 TRPV2 通道中，并研究这些分子的通道调节机制在 V1 和 V2 之间是否保守。这三个目标将通过通道活性的电生理学测量、定点诱变、计算生物物理学和结构信息的结合来实现。该奖项的指导部分将使我能够获得必要的培训，以了解计算生物物理学方法并将其严格应用到我未来的研究中。这将补充我作为实验生理学家的培训，为我提供扩展的概念框架和一套实用工具，以在原子尺度的结构动力学水平上研究蛋白质功能。我认为蛋白质结构、功能和动力学的整合对于充分理解控制蛋白质变构的机制是必要的，因此我认为我将获得的培训将是我职业生涯的基础。此外，在 Swartz 实验室（全球离子通道研究领域领先的实验室之一）接受培训将增强我在电生理学方面的专业知识并拓宽我的整体科学视野。