Development of an automated system to measure ion channel currents using a surface-modified gold probe.
Development of an automated system to measure ion channel currents using a surface-modified gold probe.
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DOI：
10.1038/s41598-021-97237-z
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发表时间：
2021-09-09
影响因子：
4.6
通讯作者：
Ide T
中科院分区：
文献类型：
--
作者：
Hirano M;Tomita M;Takahashi C;Kawashima N;Ide T
人工脂质双层单通道记录技术已用于确定各种离子通道的生物物理和药理学特性。然而，其测量效率非常低，因为它需要两个耗时的过程：脂质双层膜的制备和离子通道掺入膜中。为了解决这些问题，我们之前开发了一种基于亲水修饰金探针的技术，其上固定有离子通道，在探针亲水区域形成膜的同时，离子通道可以迅速融入双层膜中。在这里，我们通过优化金探针进一步改进了这项技术，并开发了自动通道电流测量系统。我们发现，使用圆形尖端的探针提高了通道电流测量的效率，并且除了亲水区域之外，在探针表面引入疏水区域，通过提高膜稳定性进一步提高了测量效率。此外，我们还开发了使用优化探针的自动测量系统；它使我们能够自动测量通道电流并分析阻断剂对通道活动的影响。我们的研究将有助于高通量设备的开发，以识别影响离子通道活性的候选药物。
Artificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and incorporation of ion channels into the membranes. In order to address these problems, we previously developed a technique based on hydrophilically modified gold probes on which are immobilized ion channels that can be promptly incorporated into the bilayer membrane at the same time as the membrane is formed on the probes’ hydrophilic area. Here, we improved further this technique by optimizing the gold probe and developed an automated channel current measurement system. We found that use of probes with rounded tips enhanced the efficiency of channel current measurements, and introducing a hydrophobic area on the probe surface, beside the hydrophilic one, further increased measurement efficiency by boosting membrane stability. Moreover, we developed an automated measurement system using the optimized probes; it enabled us to automatically measure channel currents and analyze the effects of a blocker on channel activity. Our study will contribute to the development of high-throughput devices to identify drug candidates affecting ion channel activity.