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	A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor
	Hui, X; Gao, J; Xie, X; Suto, N; Ogiku, T; Wang, MW
刊名	Acta pharmacologica sinica
	2005-10-01
卷号	26期号:10页码:1175-1180
关键词	Nicotinic acetylcholine receptor Scintillation proximity assay High-throughput screening
ISSN号	1671-4083
DOI	10.1111/j.1745-7254.2005.00202.x
通讯作者	Wang, mw()
英文摘要	Aim: to develop a homogeneous high-throughput screening (hts) assay based on scintillation proximity assay (spa) technology for identification of novel alpha 4 beta 2 nicotinic acetylcholine receptor (nachr) modulators. methods: membrane preparation of hek293 cells expressing alpha 4 beta 2 nachr, [h-3]cytisine and wheat germ agglutinin (wga)-coupled microbeads were used to develop an hts assay based on spa technology. this method was validated against a conventional filter binding approach and applied to large-scale screening of a library containing 32 000 synthetic compounds. intracellular calcium measurement was carried out to verify the bioactivities of the hits found by the spa assay. results: ic50 values of 2 reference compounds (epibatidine and rjr 2403) determined by spa and filter binding methods were comparable and consistent with those reported elsewhere. a total of 54 compounds, showing more than 60% competitive inhibition on [h-3]cytisine binding to alpha 4 beta 2 nachr, were identified initially following an hts campaign. secondary screening confirmed that 17 compounds with novel chemical structures possessed relatively high binding affinity to alpha 4 beta 2 nachr (k-i < 2 mu mol/l). eight compounds displayed antagonistic effects with > 50% inhibition on abt-594-induced calcium mobilization while none showed any agonist activity. conclusions: this homogeneous binding assay is a highly efficient, amenable to automation and robust tool to screen potential alpha 4 beta 2 nachr modulators in an hts setting. its application may be expanded to other membrane receptors and ion channels.
WOS关键词	SCINTILLATION-PROXIMITY ASSAYS ; ABT-594 ; PHARMACOLOGY ; CELLS
WOS研究方向	Chemistry ; Pharmacology & Pharmacy
WOS类目	Chemistry, Multidisciplinary ; Pharmacology & Pharmacy
语种	英语
出版者	BLACKWELL PUBLISHING
WOS记录号	WOS:000232346000004
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2378146
专题	中国科学院大学
通讯作者	Wang, MW
作者单位	1.Chinese Acad Sci, Grad Sch, Chinese Acad Sci,Shanghai Inst Mat Med, Shanghai Inst Biol Sci,Nat Ctr Drug Screening, Shanghai 201203, Peoples R China 2.Tanabe Seiyaku Co Ltd, Discovery & Pharmacol Res Labs, Toda, Saitama 3358505, Japan
推荐引用方式 GB/T 7714	Hui, X,Gao, J,Xie, X,et al. A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor[J]. Acta pharmacologica sinica,2005,26(10):1175-1180.
APA	Hui, X,Gao, J,Xie, X,Suto, N,Ogiku, T,&Wang, MW.(2005).A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor.Acta pharmacologica sinica,26(10),1175-1180.
MLA	Hui, X,et al."A robust homogeneous binding assay for alpha 4 beta 2 nicotinic acetylcholine receptor".Acta pharmacologica sinica 26.10(2005):1175-1180.

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