Mechanically stable solvent-free lipid bilayers in nano- and micro-tapered apertures for reconstitution of cell-free synthesized hERG channels

Daisuke Tadaki; Daichi Yamaura; Shun Araki; Miyu Yoshida; Kohei Arata; Takeshi Ohori; Ken-Ichi Ishibashi; Miki Kato; Teng Ma; Ryusuke Miyata; Yuzuru Tozawa; Hideaki Yamamoto; Michio Niwano; Ayumi Hirano-Iwata

doi:10.1038/s41598-017-17905-x

Mechanically stable solvent-free lipid bilayers in nano- and micro-tapered apertures for reconstitution of cell-free synthesized hERG channels

Sci Rep. 2017 Dec 18;7(1):17736. doi: 10.1038/s41598-017-17905-x.

Authors

Affiliations

¹ Laboratory for Nanoelectronics and Spintronics, Research Institute of Electrical Communication, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, 980-8577, Japan.
² Hang-Ichi Corporation, 1-7-315 Honcho, Naka-ku, Yokohama, Kanagawa, 231-0005, Japan.
³ Advanced Institute for Materials Research, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, 980-8577, Japan.
⁴ Department of Chemistry, Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama, Saitama, 338-8570, Japan.
⁵ Frontier Research Institute for Interdisciplinary Sciences, Tohoku University, 6-3 Aramaki-Aza-Aoba, Aoba-ku, Sendai, Miyagi, 980-8578, Japan.
⁶ Kansei Fukushi Research Institute, Tohoku Fukushi University, 6-149-1 Kunimi-ga-oka, Aoba-ku, Sendai, Miyagi, 989-3201, Japan.
⁷ Laboratory for Nanoelectronics and Spintronics, Research Institute of Electrical Communication, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, 980-8577, Japan. [email protected].
⁸ Advanced Institute for Materials Research, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi, 980-8577, Japan. [email protected].

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Membrane / metabolism
Ether-A-Go-Go Potassium Channels / metabolism*
Ether-A-Go-Go Potassium Channels / physiology
Humans
Ion Channels / metabolism
Lipid Bilayers / chemical synthesis
Lipid Bilayers / metabolism*
Micro-Electrical-Mechanical Systems / methods*
Microtechnology / methods
Potassium Channels / metabolism
Potassium Channels / physiology
Silicon Compounds
Solvents

Substances

Ether-A-Go-Go Potassium Channels
Ion Channels
KCNH1 protein, human
Lipid Bilayers
Potassium Channels
Silicon Compounds
Solvents
silicon nitride