A Comparison of the Effects of Substrate Activity on Cytochrome P450 Enzymes

Cytochromes P450 (CYP-450) are a superfamily of enzymes that are mostly found in the liver and whose job is vital in metabolizing drugs and xenobiotics, as well as being involved in the synthesis of steroids and vitamins. Some of these enzymes are substrate specific while others can work with an array of substrates. In order to determine substrate specificity for the wild-type and the enzyme with one mutation E267V, we performed enzyme assays, with the purpose of comparing enzyme activity. The two substrates of interest are N-palmitoyl glycine (NPG) and N-dimethylbenzylamine. In addition to the enzymatic assays, we determined the equilibrium association (Ka) and equilibrium dissociation (Kd) constants in order to understand the binding affinity of the enzyme and substrate to create the enzyme-substrate complex. We hypothesize that NPG will have a greater affinity to bind to the enzyme and show increased enzymatic activity for the wild-type enzyme.