Structure and function of Transient Receptor Potential channels

项目摘要 瞬时受体电位（TRP）通道受多种刺激调节，包括化学物质， 温度，机械应力和膜电压，并直接参与感官的感知 例如视觉、味觉、嗅觉、听觉、触觉、温度和疼痛。TRP通道与 在许多疾病的发病机制中，其中一些是重要的预后标志物， 有希望的新的治疗策略，以治疗各种人类癌症的目标。为了有效的药物设计， 我们需要有关TRP通道结构和功能的详细信息。我们计划研究TRP通道 使用不同的生物物理学和生物化学方法的组合来研究结构和功能。我们的具体目标 建立钙离子调节TRP通道的分子基础， TRP通道门控机制，以及3）开发各种TRP通道抑制的分子模型 有机和无机分子。TRP通道是结构-功能研究中具有挑战性的目标 因为它们代表大尺寸的多聚体整合膜蛋白 程度.为了实现我们的目标，我们将结合使用结构和功能方法，包括 现代晶体学技术，基于双折射的尺寸排阻色谱法（FSEC），钙 成像、荧光光谱和电生理学。我们将使用不同的结晶方法， 温度，筛选洗涤剂，脂质和配体，以获得不同细胞中完整的TRP通道的结构。 构象状态然后，我们将联合收割机将新生结构信息与功能数据相结合， TRP通道调控和门控机制。实现我们的目标将对 感觉生物学，并将导致一个新的结构/功能模型的TRP通道，可以作为一个 用于理论预测、计算机拟合和化学合成新药的动态模板。

PROJECT SUMMARY Transient Receptor Potential (TRP) channels are regulated by a broad range of stimuli, including chemicals, temperature, mechanical stress and membrane voltage and are directly involved in the perception of sensory modalities such as vision, taste, olfaction, hearing, touch, temperature and pain. TRP channels are implicated in the pathogenesis of numerous diseases and some of them represent important prognostic markers and promising targets for new therapeutic strategies to treat a variety of human cancers. For efficient drug design, we need detailed information about TRP channel structure and function. We plan to study TRP channel structure and function using a combination of different biophysical and biochemical methods. Our specific aims are: 1) establish molecular bases of TRP channel regulation by calcium, 2) determine the molecular mechanism of TRP channel gating, and 3) develop molecular models of TRP cannel inhibition by various organic and inorganic molecules. TRP channels are challenging targets for structure-functional studies because they represent multimeric integral membrane proteins of a large size with typically low expression levels. To achieve our goals, we will use a combination of structural and functional approaches including modern crystallographic techniques, Fluorescence-based Size Exclusion Chromatography (FSEC), calcium imaging, fluorescent spectroscopy and electrophysiology. We will use different crystallization methods and temperatures, screen detergents, lipids and ligands to obtain structures of intact TRP channels in different conformational states. We will then combine the nascent structural information with functional data to discern mechanisms of TRP channel regulation and gating. Achieving our aims will have a significant impact on sensory biology and will result in a new structural/functional model of TRP channel that can serve as a dynamic template for theoretical prediction, in silico fitting and chemical synthesis of new drugs.