Ultrasensitive Liposome-Based Assay for the Quantification of Fundamental Ion Channel Properties,Analytica Chimica Acta

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Ultrasensitive Liposome-Based Assay for the Quantification of Fundamental Ion Channel Properties
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-05-01 , DOI: 10.1016/j.aca.2020.03.044
Yi Shen ₁ , Yulong Zhong ₂ , Fan Fei ₁ , Jielin Sun ₃ , Daniel M Czajkowsky ₁ , Bing Gong ₂ , Zhifeng Shao ₁

Affiliation

One of the most widely used approaches to characterize transmembrane ion transport through nanoscale synthetic or biological channels is a straightforward, liposome-based assay that monitors changes in ionic flux across the vesicle membrane using pH- or ion-sensitive dyes. However, failure to account for the precise experimental conditions, in particular the complete ionic composition on either side of the membrane and the inherent permeability of ions through the lipid bilayer itself, can prevent quantifications and lead to fundamentally incorrect conclusions. Here we present a quantitative model for this assay based on the Goldman-Hodgkin-Katz flux theory, which enables accurate measurements and identification of optimal conditions for the determination of ion channel permeability and selectivity. Based on our model, the detection sensitivity of channel permeability is improved by two orders of magnitude over the commonly used experimental conditions. Further, rather than obtaining qualitative preferences of ion selectivity as is typical, we determine quantitative values of these parameters under rigorously controlled conditions even when the experimental results would otherwise imply (without our model) incorrect behavior. We anticipate that this simply employed ultrasensitive assay will find wide application in the quantitative characterization of synthetic or biological ion channels.

中文翻译：

用于定量基本离子通道特性的超灵敏脂质体检测

表征通过纳米级合成或生物通道的跨膜离子传输的最广泛使用的方法之一是一种简单的基于脂质体的测定，它使用 pH 或离子敏感染料监测跨囊泡膜的离子通量变化。然而，未能考虑到精确的实验条件，特别是膜两侧的完整离子组成和离子通过脂质双层本身的固有渗透性，可能会阻止量化并导致根本不正确的结论。在这里，我们基于 Goldman-Hodgkin-Katz 通量理论提出了该测定的定量模型，该模型能够准确测量和确定用于确定离子通道渗透性和选择性的最佳条件。基于我们的模型，通道渗透率的检测灵敏度比常用的实验条件提高了两个数量级。此外，我们不是像典型的那样获得离子选择性的定性偏好，而是在严格控制的条件下确定这些参数的定量值，即使实验结果可能暗示（没有我们的模型）不正确的行为。我们预计这种简单使用的超灵敏检测将在合成或生物离子通道的定量表征中得到广泛应用。我们在严格控制的条件下确定这些参数的定量值，即使实验结果会暗示（没有我们的模型）不正确的行为。我们预计这种简单使用的超灵敏检测将在合成或生物离子通道的定量表征中得到广泛应用。我们在严格控制的条件下确定这些参数的定量值，即使实验结果会暗示（没有我们的模型）不正确的行为。我们预计这种简单使用的超灵敏检测将在合成或生物离子通道的定量表征中得到广泛应用。

更新日期：2020-05-01

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