Abstract
Cytochrome P450cam (CYP101) is a monooxygenase found in the soil bacterium Pseudomonas putida. In the presence of its electron transfer partner putidaredoxin (Pdx), CYP101 catalyzes the reaction of 5-exo hydroxylation of camphor. With a significant number of backbone resonances already assigned in this large protein in its substrate camphor- and CO-bound reduced form, we investigated the binding effect between mutant CYP101 and Pdx. Nuclear magnetic resonance (NMR) spectroscopy showed that the binding between these two proteins decreased, compared to the wild type (WT) CYP101 and Pdx. Cytochrome P450 3A4 (CYP3A4), as a member of the cytochrome P450 oxidase family, is one of the most important enzymes in the human body. It is involved in the metabolism of a large number of xenobiotics. Solution state NMR study on this membrane bound protein was accomplished by the incorporation of CYP3A4 into nanodiscs, which prevents CYP3A4 from aggregating and keeps CYP3A4 as a monomer.