ISSN: 1920-4159
+44 1300 500008
Wenhui Liu
China
Short Communication
Coexpression of Genetically Engineered Cyt b5-CYP3A4 Fusion Protein with
POR in Sf9 Insect Cells and Functional Characterization of the Expressed
Products in vitro
Author(s): Zhangming Xie, Shabbir Ahmed, Wenhui Liu, Sisi Kong, Yingchun Xu, Ting Liu and Shuqing Chen
Zhangming Xie, Shabbir Ahmed, Wenhui Liu, Sisi Kong, Yingchun Xu, Ting Liu and Shuqing Chen
Human cytochrome P450 3A4 (CYP3A4) is the most abundant phase I drug-metabolizing enzyme in the liver, and approximately 50% of drugs on the market are metabolized by CYP3A4. Therefore, many in vitro studies relied on recombinant CYP3A4 as screening tool to evaluate potential drug-drug interactions (DDIs) in vivo. However, limited information regarding recombinant CYP3A4 with high catalytic activity is available. So, the present study aimed to obtain recombinant CYP3A4 with high catalytic activity and to characterize its functions in vitro. To enhance the catalytic activities of heterologously expressed CYP3A4, the enzyme was fused to cytochrome b5 (b5) tail-to-head, and the fused enzyme was inserted together with NADPH–P450 reductase (POR) into a single plasmid to achieve a simultaneous expression in sf9 cells. Here, substrate binding affinities, enzymatic activities and applic.. View More»
DOI:
10.21065/1920-4159.1000223