Phospholipase A2-catalyzed conversion of hepatic cytochrome P450 to P420 in microsomal membranes prepared from young and old rats

Abstract

Previous studies from this Laboratory have revealed few significant age-related changes in either hepatic microsomal cytochrome P450 content or NADPH-cytochrome P450 reductase activity, prompting the suggestion that the nearly ubiquitous age-associated decline in the hepatic metabolism of xenobiotics by the mixed-function oxidases is a result of alterations in the lipid matrix in which the macromolecular components are organized. In order to examine this postulate, the membrane microenvironment surrounding hepatic microsomal cytochrome P450 was enzymatically digested with snake venom phospholipase A2, and the kinetics of conversion of cytochrome P450 to cytochrome P420 determined for preparations obtained from rats of varying age. There were no significant differences in the kinetics of conversion of cytochrome P450 to cytochrome P420, suggesting that there are no significant age-related changes in the lipid microenvironment immediately adjacent to cytochrome P450. However, there was a slight difference, independent of age, in the rate of conversion of cytochrome P450 to cytochrome P420 between smooth and rough microsomes. The results are discussed in terms of general experimental approaches toward understanding drug metabolism in senescent organisms.