The cytochromes P-450 metabolize exogenous materials such as carcinogens and drugs as well as endogenous ones such as cholesterol, fatty acids and prostaglandins. The focus of this project is the purification and characterization of these isozymes. Many forms of P-450 were purified from rat liver and kidney, and rabbit liver microsomes. These purified P-450s were characterized catalytically, structurally and spectrally, and classified on the basis of immunological cross-reactivity. Several forms of P-450 were used to study the conformation of the protein surface, using monoclonal antibodies (MAbs) to specific cytochromes P-450. MAbs were also used for immunopurification of P-450s with catalytic activity, for making a P-450-gioreactor and for evaluating epitope topography of the microsomal membrane.
