Expression, Purification and Characterization of CAR/NCOA-1 Tethered Protein in <i>E. coli</i> Using Maltose-Binding Protein Fusion Tag and Gelatinized Corn Starch

The constitutive active/androstane receptor (CAR) is a nuclear that functions as xenobiotic sensor, which regulates the expression of enzymes involved in drug metabolism and efflux transporters. Evaluation binding properties between CAR was assumed to facilitate prediction drug–drug interaction, thereby contributing discovery. purpose this study construct system for rapid evaluation interactions drugs. We prepared recombinant protein using Escherichia coli system. Since isolated known be unstable, we designed fusion with sequence coactivator 1 (NCOA1), expressed maltose (MBP), purified it by several chromatography steps. thus-obtained CAR/NCOA1 tethered (CAR-NCOA1) used evaluate agonists inverse thermal denaturation experiment differential scanning fluorometry (DSF) presence absence An increase melting temperature observed addition drugs, confirming direct interaction them CAR. DSF easy set up compatible multiwell plate devices (such 96-well plates). use CAR-NCOA1 together allows CAR, considered useful