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 app...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.展开更多
文摘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.
