Structure and function of Transient Receptor Potential channels

项目总结 瞬时受体电位(Trp)通道受多种刺激的调节，包括化学物质， 温度、机械应力和膜电压直接参与感官知觉 例如视觉、味觉、嗅觉、听觉、触觉、温度和疼痛。牵涉到TRP通道 在许多疾病的发病机制中，其中一些是重要的预后标志和 有希望成为治疗多种人类癌症的新治疗策略的目标。为了高效的药物设计， 我们需要关于Trp通道结构和功能的详细信息。我们计划研究Trp渠道 利用不同的生物物理和生化方法相结合的结构和功能。我们的具体目标 1)建立钙调节Trp通道的分子基础；2)确定分子 Trp通道的门控机制；3)建立了不同途径抑制Trp通道的分子模型。 有机和无机分子。Trp通道是结构-功能研究的具有挑战性的靶点 因为它们代表的是通常低表达的大尺寸多聚体完整膜蛋白 级别。为了实现我们的目标，我们将使用结构和功能相结合的方法，包括 现代结晶学技术，基于荧光的尺寸排除层析(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和 温度、洗涤剂、脂类和配体的筛选以获得不同环境中完整的Trp通道的结构 构象状态。然后，我们将结合新出现的结构信息和功能数据来识别 色氨酸通道调节和门控的机制。实现我们的目标将对……产生重大影响 并将导致一种新的色氨酸通道结构/功能模型，它可以作为一种 用于理论预测的动态模板，用于硅胶拟合和新药的化学合成。

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.