Regular ArticleExpression, Purification, and Characterization of a Catalytically Active Human Cytochrome P450 1A2:Rat NADPH-Cytochrome P450 Reductase Fusion Protein☆
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2011, Protein Expression and PurificationCitation Excerpt :This result suggests that the tricistronic expression vector in E. coli was effective for the simultaneous expression of the three components between 24 and 48 h following incubation. It has been reported elsewhere that P450 expression can be modulated by co-expression together with other proteins, and its content levels are higher when the P450 is expressed monocistronically [35]. Using the multiple expression method with the tricistronic vector, it was possible to express P450 27A1 to a level of 330 nmol/L of culture medium when coexpressed with Adx and Adr in the tricistronic system (as judged by spectral analysis after 48 h, Supplementary Data Fig. S-1), which we considered adequate for further studies (the level was 300 nmol/L in a separate experiment).
Drug metabolism by CYP2C8.3 is determined by substrate dependent interactions with cytochrome P450 reductase and cytochrome b5
2011, Biochemical PharmacologyCitation Excerpt :For human liver microsomes, NADPH-cytochrome c reduction activities were determined as described previously [35]. P450 reductase concentrations were calculated assuming a specific activity of 3.0 μmol of cytochrome c reduced per minute per nanomole of reductase, based on published data for purified human and rabbit reductase preparations [15]. The concentration of cytochrome b5 was calculated spectrally following published methods [28,36].
High-level expression of human cytochrome P450 1A2 by co-expression with human molecular chaperone HDJ-1(Hsp40)
2004, Protein Expression and Purification