Coexpression of PXRLBD with SRC88 and construction of equilibrium dialysis model of screening PXR ligands.
- Author:
Shan-Shan YE
1
;
Chun-Na YU
;
Jing CHEN
;
Hong-Ying SUN
;
Shu-Qing CHEN
Author Information
1. Institute of Pharmacology and Toxicology and Biochemical Pharmacy, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
- Publication Type:Journal Article
- MeSH:
Clotrimazole;
metabolism;
Dexamethasone;
metabolism;
Dialysis;
methods;
Drug Interactions;
Escherichia coli;
genetics;
metabolism;
Histone Acetyltransferases;
genetics;
metabolism;
Humans;
Ligands;
Nuclear Receptor Coactivator 1;
Plasmids;
Protein Binding;
Receptors, Steroid;
genetics;
metabolism;
Transcription Factors;
genetics;
metabolism;
Transformation, Genetic
- From:
Acta Pharmaceutica Sinica
2008;43(4):427-430
- CountryChina
- Language:Chinese
-
Abstract:
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.
