Asakura Mami, Wang Shuyan, Hirano Minako, Ide Toru
Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, 3-1-1 Tsushima-naka, Kita-ku, Okayama-shi, Okayama, 700-8530, Japan.
Department of Comprehensive Technical Solutions, Okayama University, 3-1-1 Tsushima-naka, Kita-ku, Okayama-shi, Okayama, 700-8530, Japan.
Anal Sci. 2025 Jul;41(7):1073-1082. doi: 10.1007/s44211-025-00792-y. Epub 2025 May 20.
In this paper, we report the development of a device that improves the conventional artificial lipid bilayer method and can measure channel currents more efficiently. Ion channel proteins are an attractive research target in biophysics, because their functions can be measured at the single-molecule level with high time resolution. In addition, they have attracted attention as targets for drug discovery because of their crucial roles in vivo. Although electrophysiological methods are powerful tools for studying channel proteins, they suffer from low measurement efficiency and require considerable skill. In our previous paper, we reported that by immobilizing channel proteins on agarose gel beads and forming an artificial lipid bilayer on the bead surface, we simultaneously solved two problems that had been hindering the efficiency of the artificial bilayer method: the time-consuming formation of artificial lipid bilayers and the time-consuming incorporation of channels into artificial bilayers. Previous studies have utilized crosslinked hard beads; however, here we show that channel current measurement can be achieved more simply and efficiently using non-crosslinked soft beads. In this study, we detailed the process of immobilizing channel proteins on the surface of non-crosslinked beads through chemical modification, allowing us to measure their channel activity. This method enables current measurements without the need for stringent bead size selection or high negative pressure.
在本文中，我们报告了一种装置的开发，该装置改进了传统的人工脂质双层方法，能够更高效地测量通道电流。离子通道蛋白是生物物理学中一个有吸引力的研究目标，因为其功能可以在单分子水平上以高时间分辨率进行测量。此外，由于它们在体内的关键作用，它们作为药物发现的靶点也受到了关注。尽管电生理方法是研究通道蛋白的有力工具，但它们存在测量效率低的问题，并且需要相当的技巧。在我们之前的论文中，我们报告说，通过将通道蛋白固定在琼脂糖凝胶珠上，并在珠表面形成人工脂质双层，我们同时解决了一直阻碍人工双层方法效率的两个问题：人工脂质双层形成耗时以及通道掺入人工双层耗时。先前的研究使用了交联硬珠；然而，在这里我们表明，使用非交联软珠可以更简单有效地实现通道电流测量。在本研究中，我们详细阐述了通过化学修饰将通道蛋白固定在非交联珠表面的过程，从而能够测量它们的通道活性。这种方法无需严格选择珠子大小或施加高负压就能进行电流测量。
