Electrical Spiking in Escherichia coli Probed with a Fluorescent Voltage-Indicating Protein
Electrical Spiking in Escherichia coli Probed with a Fluorescent Voltage-Indicating Protein
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DOI：
10.1126/science.1204763
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发表时间：
2011-07-15
期刊：
影响因子：
56.9
通讯作者：
Cohen, Adam E.
中科院分区：
文献类型：
--
作者：
Kralj, Joel M.;Hochbaum, Daniel R.;Cohen, Adam E.
细菌有许多电压门控和配体门控离子通道，群体水平的测量表明，膜电位对细菌的生存很重要。然而，在完整的细菌中探测电压动态是不可能的。在这里，我们开发了一种方法来揭示大肠杆菌中的电尖峰。为了探测细菌膜电位，我们设计了一种基于绿色吸收蛋白视紫红质的电压敏感荧光蛋白。蛋白视紫红质光学质子传感器（PROPS）在大肠杆菌中的表达。大肠杆菌显示了高达1赫兹的电尖峰。尖峰是敏感的化学和物理扰动，并与快速流出的小分子荧光团，这表明细菌外排机制可能是电调节。
Bacteria have many voltage-and ligand-gated ion channels, and population-level measurements indicate that membrane potential is important for bacterial survival. However, it has not been possible to probe voltage dynamics in an intact bacterium. Here we developed a method to reveal electrical spiking in Escherichia coli. To probe bacterial membrane potential, we engineered a voltage-sensitive fluorescent protein based on green-absorbing proteorhodopsin. Expression of the proteorhodopsin optical proton sensor (PROPS) in E. coli revealed electrical spiking at up to 1 hertz. Spiking was sensitive to chemical and physical perturbations and coincided with rapid efflux of a small-molecule fluorophore, suggesting that bacterial efflux machinery may be electrically regulated.