Cytochrome P-450 reductase FAD binding domain: Cloning, enzymatic activity and flavin binding characterization
Abstract
The Mixed Function Oxidase System metabolizes a wide range of biochemicals including drugs, pesticides and steroids. Cytochrome P450 reductase is a key enzymatic component of this system, supplying reducing equivalents from NADPH to cytochrome P450. The electrons are shuttled through reductase via two flavin moieties: FAD and FMN. Although the exact mechanism of flavins action is not known, the enzymatic features of reductase greatly depleted of either FMN of FAD have been characterized. Additionally, flavin location within reductase has been proposed by homology and chemical modification studies. This study seeks to extend the flavin depletion analysis in a more controlled system by eliminating the proposed FMN binding domain with recombinant DNA techniques and biochemical analysis. Two P450 reductase cDNA clones containing only the FMN and NADPH binding domain were isolated, expressed and the protein products purified and analysed. This study confirms the proposed FAD binding site, role of FAD in electron shuttling pathway and provides new methods to study the FAD binding domain.
Subject Area
Biochemistry|Molecular biology
Recommended Citation
Hodgson, Anne Veronica, "Cytochrome P-450 reductase FAD binding domain: Cloning, enzymatic activity and flavin binding characterization" (1994). Texas Medical Center Dissertations (via ProQuest). AAI9520977.
https://digitalcommons.library.tmc.edu/dissertations/AAI9520977