Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, United Kingdom.
J Am Chem Soc. 2011 Jun 22;133(24):9370-5. doi: 10.1021/ja200128n. Epub 2011 May 27.
The ability to routinely study eukaryotic ion channels in a synthetic lipid environment would have a major impact on our understanding of how different lipids influence ion channel function. Here, we describe a straightforward, detergent-free method for the in vitro reconstitution of eukaryotic ion channels and ionotropic receptors into droplet interface bilayers and measure their electrical activity at both the macroscopic and single-channel level. We explore the general applicability of this method by reconstitution of channels from a wide range of sources including recombinant cell lines and native tissues, as well as preparations that are difficult to study by conventional methods including erythrocytes and mitochondria.
如果能够在合成脂质环境中常规研究真核离子通道,将会极大地促进我们对不同脂质如何影响离子通道功能的理解。在这里,我们描述了一种简单、无去污剂的方法,用于将真核离子通道和离子型受体体外重建到液滴界面双层中,并在宏观和单通道水平上测量它们的电活性。我们通过从广泛的来源(包括重组细胞系和天然组织以及传统方法难以研究的制剂,如红细胞和线粒体)中重建通道来探索这种方法的普遍适用性。