Automated Parallel Recordings of Topologically Identified Single Ion Channels

作者:Kawano Ryuji; Tsuji Yutaro; Sato Koji; Osaki Toshihisa; Kamiya Koki; Hirano Minako; Ide Toru; Miki Norihisa; Takeuchi Shoji*
来源:Scientific Reports, 2013, 3(1): 1995.
DOI:10.1038/srep01995

摘要

Although ion channels are attractive targets for drug discovery, the systematic screening of ion channel-targeted drugs remains challenging. To facilitate automated single ion-channel recordings for the analysis of drug interactions with the intra- and extracellular domain, we have developed a parallel recording methodology using artificial cell membranes. The use of stable lipid bilayer formation in droplet chamber arrays facilitated automated, parallel, single-channel recording from reconstituted native and mutated ion channels. Using this system, several types of ion channels, including mutated forms, were characterised by determining the protein orientation. In addition, we provide evidence that both intra- and extracellular amyloid-beta fragments directly inhibit the channel open probability of the hBK channel. This automated methodology provides a high-throughput drug screening system for the targeting of ion channels and a data-intensive analysis technique for studying ion channel gating mechanisms.

  • 出版日期2013-6-17