共表达PXRLBD和SRC88以及PXR配体筛选的平衡透析模型的建立  

Coexpression of PXRLBD with SRC88 and construction of equilibrium dialysis model of screening PXR ligands

作　　者：叶姗姗[1] 俞春娜[1] 陈静[1] 孙红颖[1] 陈枢青[1] 

机构地区：[1]浙江大学药理毒理与生化药学研究所,浙江杭州310058

出　　处：《药学学报》2008年第4期427-430,共4页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30472171).

摘　　要：The aim of this study was to obtain the soluble protein of human pregnane X receptor ligand binding domain(PXRLBD) through the coexpression of PXRLBD and 88 amino acids of steroid receptor coactivator-1(SRC88) and apply the protein to constructing a new model of screening PXR ligands.Expression plasmid of pETDuet-1-SRC88-PXRLBD was constructed and transformed into Escherichia coli Rosetta(DE3) to coexpress PXRLBD and SRC88 via induction by IPTG at low temperature.Then an equilibrium dialysis model was constructed to study the interaction between PXRLBD and drugs including clotrimazole and dexamethasone,using HPLC as the analysis method.The results showed that the soluble protein of PXRLBD was obtained and the HPLC data indicated that clotrimazole bound to PXRLBD,while dexamethasone did not bind to PXRLBD,which indicated the successful establishment of a new method for studying the interaction between PXR and drugs.The new method may be useful in the screening of PXR ligands in vitro.The aim of this study was to obtain the soluble protein of human pregnane X receptor ligand binding domain （PXRLBD） through the coexpression of PXRLBD and 88 amino acids of steroid receptor coactivator-1 （SRC88） and apply the protein to constructing a new model of screening PXR ligands. Expression plasmid of pETDuet-l-SRC88-PXRLBD was constructed and transformed into Escherichia coli Rosetta （DE3） to coexpress PXRLBD and SRC88 via induction by IPTG at low temperature. Then an equilibrium dialysis model was constructed to study the interaction between PXRLBD and drugs including clotrimazole and dexamethasone, using HPLC as the analysis method. The results showed that the soluble protein of PXRLBD was obtained and the HPLC data indicated that clotrimazole bound to PXRLBD, while dexamethasone did not bind to PXRLBD, which indicated the successful establishment of a new method for studying the interaction between PXR and drugs. The new method may be useful in the screening of PXR ligands in vitro.

关 键 词：孕烷X受体 甾体受体辅活化因子 共表达 配体 透析 高效液相色谱法 

分 类 号：R963[医药卫生—微生物与生化药学]