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首页> 外文期刊>Biophysical Journal >Transient potential gradients and impedance measures of tethered bilayer lipid membranes: Pore-forming peptide insertion and the effect of electroporation
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Transient potential gradients and impedance measures of tethered bilayer lipid membranes: Pore-forming peptide insertion and the effect of electroporation

机译：拴系双层脂质膜的瞬态电位梯度和阻抗测量：成孔肽插入和电穿孔的影响

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摘要

In this work, we present experimental data, supported by a quantitative model, on the generation and effect of potential gradients across a tethered bilayer lipid membrane (tBLM) with, to the best of our knowledge, novel architecture. A challenge to generating potential gradients across tBLMs arises from the tethering coordination chemistry requiring an inert metal such as gold, resulting in any externally applied voltage source being capacitively coupled to the tBLM. This in turn causes any potential across the tBLM assembly to decay to zero in milliseconds to seconds, depending on the level of membrane conductance. Transient voltages applied to tBLMs by pulsed or ramped direct-current amperometry can, however, provide current-voltage (I/V) data that may be used to measure the voltage dependency of the membrane conductance. We show that potential gradients >～150 mV induce membrane defects that permit the insertion of pore-forming peptides. Further, we report here the novel (to our knowledge) use of real-time modeling of conventional low-voltage alternating-current impedance spectroscopy to identify whether the conduction arising from the insertion of a polypeptide is uniform or heterogeneous on scales of nanometers to micrometers across the membrane. The utility of this tBLM architecture and these techniques is demonstrated by characterizing the resulting conduction properties of the antimicrobial peptide PGLa.
机译：在这项工作中，我们提供了定量模型支持的实验数据，这些数据具有跨束缚的双层脂质膜（tBLM）的电位梯度的产生和影响，据我们所知，该结构新颖。在束缚配位化学过程中，跨tBLM产生电位梯度的挑战来自于需要惰性金属（例如金）的束缚配位化学反应，从而导致任何外部施加的电压源均与tBLM电容耦合。反过来，这会导致跨tBLM组件的任何电位在几毫秒到几秒钟内衰减到零，具体取决于膜电导的水平。但是，通过脉冲或倾斜直流电流分析法施加到tBLM的瞬态电压可以提供可用于测量膜电导的电压依赖性的电流-电压（I / V）数据。我们发现电位梯度>〜150 mV会诱发膜缺陷，从而允许插入成孔肽。此外，我们在此报告了（据我们所知）新颖的使用常规低压交流阻抗谱的实时建模来确定从多肽插入产生的传导在纳米到微米尺度上是均匀还是异质的跨膜。该tBLM体系结构和这些技术的实用性通过表征抗菌肽PGLa的最终传导特性得到了证明。

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