Identification of novel KCNQ4 openers by a high-throughput fluorescence-based thallium flux assay

doi:10.1016/j.ab.2011.06.040

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	Identification of novel KCNQ4 openers by a high-throughput fluorescence-based thallium flux assay
	Li, Qunyi 2,3; Rottlander, Mario 1; Xu, Mingkai 2,3; Christoffersen, Claus Tornby 1; Frederiksen, Kristen 1; Wang, Ming-Wei2,3 ; Jensen, Henrik Sindal 1
刊名	ANALYTICAL BIOCHEMISTRY
	2011-11-01
卷号	418 期号:1 页码:66-72
关键词	KCNQ Kv7 Patch clamp Stable cell line Thallium flux (86)Rubidium flux Potassium channel opener High-throughput screening
ISSN号	0003-2697
DOI	10.1016/j.ab.2011.06.040
文献子类	Article
英文摘要	To develop a real-time thallium flux assay for high-throughput screening (HTS) of human KCNQ4 (Kv7.4) potassium channel openers, we used CHO-K1 cells stably expressing human KCNQ4 channel protein and a thallium-sensitive dye based on the permeability of thallium through potassium channels. The electrophysiological and pharmacological properties of the cell line expressing the KCNQ4 protein were found to be in agreement with that reported elsewhere. The EC50 values of the positive control compound (retigabine) determined by the thallium and (86)rubidium flux assays were comparable to and consistent with those documented in the literature. Signal-to-background (S/B) ratio and Z factor of the thallium influx assay system were assessed to be 8.82 and 0.63, respectively. In a large-scale screening of 98,960 synthetic and natural compounds using the thallium influx assay, 76 compounds displayed consistent KCNQ4 activation, and of these 6 compounds demonstrated EC50 values of less than 20 mu mol/L. and 2 demonstrated EC50 values of less than 1 mu mol/L. Taken together, the fluorescence-based thallium flux assay is a highly efficient, automatable, and robust tool to screen potential KCNQ4 openers. This approach may also be expanded to identify and evaluate potential modulators of other potassium channels. (C) 2011 Elsevier Inc. All rights reserved.
资助项目	H. Lundbeck[00000000] ; Ministry of Science and Technology of China[2009ZX09302-001]
WOS关键词	CA2+-ACTIVATED K+ CHANNELS ; HAIR-CELLS ; INNER-EAR ; ACTIVATION ; RETIGABINE ; MUTATION ; VALIDATION ; BMS-204352 ; MODULATOR ; DEAFNESS
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000294836300010
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/278363]
专题	国家新药筛选中心中科院受体结构与功能重点实验室新药研究国家重点实验室
通讯作者	Wang, Ming-Wei
作者单位	1.H Lundbeck & Co AS, Neurosci Drug Discovery, DK-2500 Copenhagen, Denmark 2.Chinese Acad Sci, Natl Ctr Drug Screening, Shanghai 201203, Peoples R China; 3.Chinese Acad Sci, State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China;
推荐引用方式 GB/T 7714	Li, Qunyi,Rottlander, Mario,Xu, Mingkai,et al. Identification of novel KCNQ4 openers by a high-throughput fluorescence-based thallium flux assay[J]. ANALYTICAL BIOCHEMISTRY,2011,418(1):66-72.
APA	Li, Qunyi.,Rottlander, Mario.,Xu, Mingkai.,Christoffersen, Claus Tornby.,Frederiksen, Kristen.,...&Jensen, Henrik Sindal.(2011).Identification of novel KCNQ4 openers by a high-throughput fluorescence-based thallium flux assay.ANALYTICAL BIOCHEMISTRY,418(1),66-72.
MLA	Li, Qunyi,et al."Identification of novel KCNQ4 openers by a high-throughput fluorescence-based thallium flux assay".ANALYTICAL BIOCHEMISTRY 418.1(2011):66-72.