| 7853177 |
Murray M, Butler AM: Identification of a reversible component in the in vitro inhibition of rat hepatic cytochrome P450 2B1 by parathion. J Pharmacol Exp Ther. 1995 Feb;272(2):639-44.
Cytochrome P450 (P450) enzymes are inactivated in suicidal fashion during microsomal parathion oxidation. In the present study, two distinct components of the inhibition of the phenobarbital (PB)-inducible P450 2B1 by parathion were characterized. Here we report for the first time that low concentrations of parathion potently and reversibly inhibited, but did not inactivate, 2B1. In contrast, the previously described inactivation process occurred only at considerably higher parathion concentrations, at which concentrations enzyme activity was already extensively inhibited. At low concentration, parathion was a competitive inhibitor of 2B1-mediated androstenedione 16 beta-hydroxylation (Ki = 0.44 +/- 0.07 microM) and of 7-pentylresorufin O-depentylation (Ki = 0.40 +/- 0.03 microM) in microsomes from PB-pretreated rats and was similarly effective against androstenedione 16 alpha- and 16 beta-hydroxylation catalyzed by purified 2B1. Although preincubation of higher concentrations of parathion (> 5 microM) with NADPH-supplemented microsomes from PB-pretreated rat liver decreased holo-P450, heme loss was not observed near the Ki values. Instead, half-maximal loss of P450 occurred at 6 microM and at 9 microM parathion in PB-pretreated microsomes and in the reconstituted system, respectively. Parathion metabolism was efficient in PB-microsomes (Km values for 4-nitrophenol and paraoxon formation were 13 microM and 10 microM, respectively) and in the reconstituted system (corresponding Km values were 19 microM and 14 microM). Thus the constants for P450 inactivation and for parathion metabolism were similar and were at least 15-fold greater than the Ki values for the reversible process.(ABSTRACT TRUNCATED AT 250 WORDS) |
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