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首页> 外文期刊>Neuron >A fluorometric approach to local electric field measurements in a voltage-gated ion channel.
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A fluorometric approach to local electric field measurements in a voltage-gated ion channel.

机译：在电压门控离子通道中进行局部电场测量的荧光方法。

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摘要

Site-specific electrostatic measurements have been limited to soluble proteins purified for in vitro spectroscopic characterization or proteins of known structure; however, comparable measurements have not been made for functional membrane bound proteins. Here, using an electrochromic fluorophore, we describe a method to monitor localized electric field changes in a voltage-gated potassium channel. By coupling the novel probe Di-1-ANEPIA to cysteines in Shaker and tracking field-induced optical changes, in vivo electrostatic measurements were recorded with submillisecond resolution. This technique reports dynamic changes in the electric field during the gating process and elucidates the electric field profile within Shaker. The extension of this method to other membrane bound proteins, including transporters, will yield insight into the role of electrical forces on protein function.
机译：特定位置的静电测量仅限于为体外光谱表征纯化的可溶性蛋白质或已知结构的蛋白质;然而，尚未对功能性膜结合蛋白进行类似的测量。在这里，我们使用电致变色荧光团，描述了一种监测电压门控钾通道中局部电场变化的方法。通过将新型探针Di-1-ANEPIA与Shaker中的半胱氨酸偶联并跟踪场诱导的光学变化，以亚毫秒级分辨率记录了体内静电测量值。该技术报告了选通过程中电场的动态变化，并阐明了振荡器中的电场分布。此方法扩展到其他膜结合蛋白，包括转运蛋白，将使人们深入了解电对蛋白功能的作用。

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