A yeast two-hybrid technology-based system for the discovery of PPAR gamma agonist and antagonist

doi:10.1016/j.ab.2004.09.004

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	A yeast two-hybrid technology-based system for the discovery of PPAR gamma agonist and antagonist
	Chen, Q; Chen, J; Sun, T; Shen, JH; Shen, X; Jiang, HL
刊名	ANALYTICAL BIOCHEMISTRY
	2004-12-15
卷号	335 期号:2 页码:253-259
关键词	peroxisome pro liferato r-activated receptor gamma (PPAR gamma) yeast two-hybrid CREP-binding protein (CBP) agonist antagonist alpha-galactosidase
ISSN号	0003-2697
DOI	10.1016/j.ab.2004.09.004
文献子类	Article
英文摘要	Peroxisome proliferator-activated receptor gamma (PPARgamma) is an important therapeutic drug target against several diseases such as diabetes, inflammation, dyslipidemia, hypertension, and cancer. Ligand binding to PPARgamma is responsible for controlling the biological functions, and developing new technology to measure ligand-PPARgamma binding is significant for both the function study of the receptor and ligand discovery. In this study, we exploited an efficient approach for the discovery of PPARgamma agonist and antagonist via a yeast two-hybrid system based on the fact that PPARgamma interacts with the coactivator CBP (CREP-binding protein) ligand-dependently. We employed the MELI reporter gene instead of the traditionally used LacZ gene to evaluate the protein-protein interactions by conducting a convenient alpha-galactosidase assay in the yeast strain AH109 with genes of PPARgamma-LBD (ligand-binding domain) and CBP N terminus introduced. With this built screening platform, the EC50 values of the PPARgamma agonists rosiglitazone, troglitazone, pioglitazone, indomethacin, 15-deoxy-Delta12,14-prostaglan din J(2) (15d-PGJ(2)) and GI262570 were investigated, and the quantitatively antagonistic effect by IC50 of the PPARgamma typical antagonist GW9662 on the rosiglitazone agonistic activity was fully examined. The reliability of this presented system evaluated by the comparable agreement of EC50 and IC50 values for the test compounds with the reported ones indicated that this yeast two-hybrid-based approach is powerful for PPARgamma agonist and antagonist screening. In addition, because this screening system is designed for use in a microliter plate format where numerous chemicals could be readily screened, it is hoped that this yeast two-hybrid screening approach may be adaptable for high-throughput settings. (C) 2004 Elsevier Inc. All rights reserved.
WOS关键词	ACTIVATED RECEPTOR-GAMMA ; IMPROVED INSULIN-SENSITIVITY ; TYPE-2 DIABETES-MELLITUS ; TRANSCRIPTIONAL COACTIVATORS ; PRO12ALA POLYMORPHISM ; ESTROGENIC ACTIVITY ; DRUG DISCOVERY ; BINDING-SITES ; LIGAND ; DIFFERENTIATION
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:000225502800010
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/273976]
专题	药理学第三研究室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药物发现与设计中心
通讯作者	Shen, X
作者单位	Chinese Acad Sci, Grad Sch,State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai Inst Biol Sci,Drug Discovery Design Ctr, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Chen, Q,Chen, J,Sun, T,et al. A yeast two-hybrid technology-based system for the discovery of PPAR gamma agonist and antagonist[J]. ANALYTICAL BIOCHEMISTRY,2004,335(2):253-259.
APA	Chen, Q,Chen, J,Sun, T,Shen, JH,Shen, X,&Jiang, HL.(2004).A yeast two-hybrid technology-based system for the discovery of PPAR gamma agonist and antagonist.ANALYTICAL BIOCHEMISTRY,335(2),253-259.
MLA	Chen, Q,et al."A yeast two-hybrid technology-based system for the discovery of PPAR gamma agonist and antagonist".ANALYTICAL BIOCHEMISTRY 335.2(2004):253-259.