Aldrin Epoxidation Catalyzed by Purified Rat‐Liver Cytochromes P‐450 and P‐448 High Selectivity for Cytochrome P‐450

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Abstract
Aldrin epoxidation was studied in monooxygenase systems reconstituted from purified rat livermicrosotnal cytochrome P‐450 or P‐448, NADPH‐cytochrome c reductase, dilauroylphosphatidylcholine and sodium Cholate. Cytochrome P‐450, purified from hepatic microsomes of phenobarbital‐treated rats, exhibited a high rate of dieldrin formation. The low enzyme activity observedin the absence of the lipid and sodium cholate was increased threefold by addition of dilauroyl‐phosphatidylcholine and was further stimulated twofold by addition of sodium cholate. The apparentKm for aldrin in the complete system was 7 ± 2 μM. SKF 525‐A, at a concentration of 250 μM, inhibited aldrin epoxidation by 65% whereas 7,8‐benzoflavone had no inhibitory effect at concentrations up to 250 μM. Addition of ethanol markedly increased epoxidase activity. The increase was three fold in the presence of 5% ethanol.When cytochrome P‐448 purified from hepatic microsomes of 3‐methylcholanthrene‐treated rats was used, a very low rate of epoxidation was observed which was less than 3% of the activity mediated by cytochrome P‐450 under similar assay conditions. Enzyme activity was independent of the lipid factor dilauroylphosphatidylcholine. The apparent Km for aldrin was 27 ± 7 μM. The modifiers of monooxygenase reactions, 7,8‐benzoflavone, SKF 525‐A and ethanol, inhibited the activity mediated by cytochrome P‐448. The I50 was 0.05, 0.2 and 800 mM, respectively.These results indicate that aldrin is a highly selective substrate for cytochrome P‐450 species present in microsomes of Phenobarbital‐treated animals and is a poor substrate for cytochrome P‐448. The two forms of aldrin epoxidase can be characterised by their turnover number, their apparent K, and their sensitivity to modifiers, like 7,8‐benzoflavone and ethanol.