Novel cell-free high-throughput screening method for pharmacological tools targeting K+ channels.
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.
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065.
Zhenwei
Z
Su
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065.
Emily C
EC
Brown
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065.
Weiwei
W
Wang
Laboratory of Molecular Neurobiology and Biophysics, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065 mackinn@rockefeller.edu.
Roderick
R
MacKinnon
D001145:Arrhythmias, Cardiac; D051638:Ether-A-Go-Go Potassium Channels; D057166:High-Throughput Screening Assays; D006801:Humans; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics
D051638:Ether-A-Go-Go Potassium Channels; D026902:Potassium Channel Blockers; D024642:Potassium Channels, Voltage-Gated
K+ channel screening; LFA; hERG safety assay; liposome flux assay