Abstract
1. The effect of the aromatic solvents toluene, xylene and ethylbenzene on microsomal membrane fluidity and anaerobic NADPH-reduction kinetics were studied.
2. The relation of membrane fluidity to the kinetics of cytochrome P450 reduction by NADPH-cytochrome P450 reductase was examined with regard to a membrane-mediated molecular organization of the multienzyme components of the monooxygenase system.
3. Membrane fluidity changes were detected with the steady-state pyrene excimer formation method and with fluoresence lifetime measurements after incubation of the microsomes with organic solvents.
4. Increase in membrane fluidity in presence of organic solvents leads to a small but significant decrease of the rate constant of the cytochrome P450 reduction kinetics and a change in the relative amplitudes of the components of the biphasic response.
5. The results support the idea of a molecular organization of cytochrome P450 in clusters. Fluidization of the microsomal membrane by organic solvents increase the cytochrome P450 cluster formation.