TY - JOUR
T1 - Novel cell-free high-throughput screening method for pharmacological tools targeting K+ channels
AU - Su, Zhenwei
AU - Brown, Emily C.
AU - Wang, Weiwei
AU - MacKinnon, Roderick
N1 - Funding Information:
We thank J. Fraser Glickman, Antonio Luz, and Jeanne Chiaravalli-Giganti at the High-Throughput and Spectroscopy Resource Center of The Rockefeller University for assistance with the screens; Maria L. Garcia and Gregory J. Kaczorowski for advice with data analyses; Stephen G. Brohawn for assistance with K2P constructs and TRAAK purification; Yi Chun Hsiung for assistance with insect and mammalian cell cultures; and members of the R.M. laboratory for helpful discussions. R.M. is an investigator in the Howard Hughes Medical Institute. Other funding support includes National Institutes of Health Grant GM43949, American Asthma Foundation extension award, The Robertson Therapeutic Development Fund, and The Bridge Fund (to R.M.).
PY - 2016/5/17
Y1 - 2016/5/17
N2 - 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.
AB - 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.
KW - HERG safety assay
KW - K channel screening
KW - LFA
KW - Liposome flux assay
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U2 - 10.1073/pnas.1602815113
DO - 10.1073/pnas.1602815113
M3 - Article
C2 - 27091997
AN - SCOPUS:84969752680
SN - 0027-8424
VL - 113
SP - 5748
EP - 5753
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 20
ER -