Su Zhenwei, Brown Emily C, Wang Weiwei, MacKinnon Roderick
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065.
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065
Proc Natl Acad Sci U S A. 2016 May 17;113(20):5748-53. doi: 10.1073/pnas.1602815113. Epub 2016 Apr 18.
K(+) channels, a superfamily of ∼80 members, control cell excitability, ion homeostasis, and many forms of cell signaling. Their malfunctions cause numerous diseases including neuronal disorders, cardiac arrhythmia, diabetes, and asthma. Here we present a novel liposome flux assay (LFA) that is applicable to most K(+) channels. It is robust, low cost, and high throughput. Using LFA, we performed small molecule screens on three different K(+) channels and identified new activators and inhibitors for biological research on channel function and for medicinal development. We further engineered a hERG (human ether-à-go-go-related gene) channel, which, when used in LFA, provides a highly sensitive (zero false negatives on 50 hERG-sensitive drugs) and highly specific (zero false positives on 50 hERG-insensitive drugs), low-cost hERG safety assay.
钾离子通道是一个由约80个成员组成的超家族，它控制细胞兴奋性、离子稳态以及多种细胞信号传导形式。其功能失调会引发多种疾病，包括神经紊乱、心律失常、糖尿病和哮喘。在此，我们展示了一种适用于大多数钾离子通道的新型脂质体通量测定法（LFA）。该方法稳健、成本低且通量高。利用LFA，我们对三种不同的钾离子通道进行了小分子筛选，鉴定出用于通道功能生物学研究和药物开发的新激活剂和抑制剂。我们进一步构建了一个hERG（人类ether-à-go-go相关基因）通道，当将其用于LFA时，可提供一种高度灵敏（对50种hERG敏感药物零假阴性）、高度特异（对50种hERG不敏感药物零假阳性）且低成本的hERG安全性测定法。
