Extraction and NMR of Cytochrome P450 Oxidoreductase

Cytochrome P450 oxidoreductase (CYPOR) is a 78 kDa membrane-bound protein in living cells. It delivers electrons to the protein cytochrome P450 in order to catalyze the metabolism of drugs and synthesis of steroids in the body. Studying CYPOR in its full length form is not an easy task. This is because most of the biophysical analyses of this protein require huge amounts of it to be extracted from the membrane. Normally, the extraction of proteins from the membrane is performed using detergents. This work states that Triton X-100 is the best detergent that extracts the CYPOR from its membrane. Furthermore, to better understand the dynamics of CYPOR, the nuclear magnetic resonance (NMR) technology is applied. This work investigates parts of this protein using NMR. It shows the effect of oxidation, reduction, and mutations on the dynamics of CYPOR.