Phospholipid bilayer membranes play decisive roles in the cytochrome P-450-dependent monooxygenase system.
Hepatic microsomal monooxygenase was reconstituted by incorporating cytochrome P-450 and NADPH-cytochrome P-450 reductase, which had been purified from phenobarbital-pretreated rabbit liver microsomes, into phospholipid liposomal membranes. The NADPH-dependent monooxygenase activity of the reconstituted system was found to be dependent on the phospholipid-to-protein ratio, i.e., the two-dimensional concentration of the two proteins on the plane of the membranes. A similar concentration dependence was also observed in the cytochrome b5 and NADH-cytochrome b5 system, which had been incorporated into liposomal membranes. The diffusion process of the proteins in the membrane, therefore, plays an important role in the monooxygenase system. When the fluidity of the membrane was changed by utilizing a synthetic dimyristoylphosphatidylcholine, which shows a well-defined gel to liquid crystalline phase transition, the activation energy of the monooxygenase reaction was changed at around the phase transition temperature, suggesting a conformational change of cytochrome P-450 caused by the fluidity change of the membrane. The incorporation of P-450 into liposomes was also found to affect the binding of substrates to cytochrome P-450. The decrease in the apparent dissociation constant of substrates upon incorporation into membranes suggests that the lipid membrane acts as a pool for hydrophobic substrates, which are concentrated in the lipid phase, and that cytochrome P-450 takes substrates directly from the membrane phase. Phospholipid membranes, therefore, play very important roles in various phases of the reaction of cytochrome P-450-dependent monooxygenase.
Taniguchi
H
H
Institute of Experimental Pathology, German Cancer Research Center, Heidelberg.
Pyerin
W
W
D000818:Animals; D003577:Cytochrome P-450 Enzyme System; D003579:Cytochrome Reductases / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism; D042966:Cytochrome-B(5) Reductase; D015786:Cytochromes b5; D008051:Lipid Bilayers / Q000378:metabolism*; D008560:Membrane Fluidity; D008862:Microsomes, Liver / Q000201:enzymology; D010105:Oxygenases / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011817:Rabbits
D003573:Cytochrome b Group; D008051:Lipid Bilayers; D010743:Phospholipids; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; D003579:Cytochrome Reductases; D042966:Cytochrome-B(5) Reductase