ION CHANNEL STRUCTURE AND FLUORESCENCE ENERGY TRANSFER

描述(摘自申请者摘要)：考察关系 电压门控离子通道的结构和功能之间的关系 阐明细胞兴奋性的基本机制，但也将 解释离子通道如何发生故障。离子通道突变已经被 与从新陈代谢紊乱到心脏等一系列疾病有关 心律失常，可能与癫痫和其他神经系统疾病有关 精神错乱。了解电压敏感性的分子机制 离子通道不仅会使我们更好地理解电学 神经元之间的交流，但也将有助于解释临床 离子通道紊乱的表现。电压门控的最新知识 离子通道表示通道的不同区域经历 构象随电压变化。然而，这些问题的规模 变化及其对渠道结构的影响是未知的，正如 蛋白质的三级结构。测量不同设备之间的距离 通道的区域作为电压的函数将有助于阐明 与电压门控相关的构象变化。荧光 利用共振能量转移(FRET)来测量距离 在两个不同的荧光探针之间，一个供体分子和一个受体 分子，通过测量探针之间的能量转移。通过 将一个探头连接到假定的电压敏感区域，而另一个连接到 通道的静态区域，FRET有可能确定 沟道区域响应于电压和距离而移动 由这些区域从通道的关闭状态行进到打开状态。

DESCRIPTION (Adapted from applicant's abstract): Examining the relationship between structure and function in voltage-gated ion channels will not only clarify the fundamental mechanisms of cellular excitability, but will also explain how ion channels can malfunction. Ion channel mutations have been linked to disorders ranging from metabolic disturbances to heart arrhythmias, with possible links to epilepsy and other neurological disorders. Understanding the molecular mechanisms of voltage sensitivity in ion channels will not only lead to a better understanding of the electrical communication between neurons, but will also help explain the clinical manifestations of ion channel disorders. Current knowledge of voltage-gated ion channels indicates that different regions of the channel undergo conformational changes with voltage. However, the magnitude of these changes and their effects on channel structure are unknown, as is the tertiary structure of the protein. Measuring distances between different regions of the channel as a function of voltage will help elucidate the conformational changes associated with voltage gating. Fluorescence resonance energy transfer (FRET) has been used to measure the distance between two different fluorescent probes, a donor molecule and an acceptor molecule, by measuring the transfer of energy between the probes. By attaching one probe to a putative voltage-sensing region and the other to a static region of the channel, FRET has the potential to determine which regions of the channel move in response to voltage and the distances traveled by these regions from the closed to the open state of the channel.