Laboratory for Nanoelectronics and Spintronics, Research Institute of Electrical Communication, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, 980-8577, Japan.
Hang-Ichi Corporation, 1-7-315 Honcho, Naka-ku, Yokohama, Kanagawa, 231-0005, Japan.
Sci Rep. 2017 Dec 18;7(1):17736. doi: 10.1038/s41598-017-17905-x.
The self-assembled bilayer lipid membrane (BLM) is the basic component of the cell membrane. The reconstitution of ion channel proteins in artificially formed BLMs represents a well-defined system for the functional analysis of ion channels and screening the effects of drugs that act on them. However, because BLMs are unstable, this limits the experimental throughput of BLM reconstitution systems. Here we report on the formation of mechanically stable solvent-free BLMs in microfabricated apertures with defined nano- and micro-tapered edge structures. The role of such nano- and micro-tapered structures on the stability of the BLMs was also investigated. Finally, this BLM system was combined with a cell-free synthesized human ether-a-go-go-related gene channel, a cardiac potassium channel whose relation to arrhythmic side effects following drug treatment is well recognized. Such stable BLMs as these, when combined with a cell-free system, represent a potential platform for screening the effects of drugs that act on various ion-channel genotypes.
自组装双层脂质膜 (BLM) 是细胞膜的基本组成部分。离子通道蛋白在人工形成的 BLM 中的再构成代表了用于离子通道功能分析和筛选作用于它们的药物效果的一种明确的系统。然而,由于 BLM 是不稳定的,这限制了 BLM 再构成系统的实验通量。在此,我们报告了在具有定义的纳米和微渐缩边缘结构的微制造孔中形成机械稳定的无溶剂 BLM。还研究了这种纳米和微渐缩结构对 BLM 稳定性的作用。最后,将该 BLM 系统与无细胞合成的人类 ether-a-go-go 相关基因通道(一种钾通道,其与药物治疗后心律失常副作用的关系已得到充分认识)相结合。当与无细胞系统结合使用时,这种稳定的 BLM 代表了筛选作用于各种离子通道基因型的药物效果的潜在平台。