Inactivation of phenobarbital-inducible rabbit-liver microsomal cytochrome P-450 by allylisopropylacetamide: impact on electron transfer.
Application of a single dose of allylisopropylacetamide (AIA) to phenobarbital-pretreated rabbits resulted in partial destruction of the heme moiety of liver microsomal cytochrome P-450. A minor fraction of chromophore loss was accounted for by heme-derived product(s) covalently attached to microsomal proteins. Interestingly, cytochrome P-450 appeared to have undergone significant drug-mediated alkylation of the apohemoprotein. The modified species was purified to apparent homogeneity and shown to arise from AIA-induced blockage of about 2 histidines in the cytochrome P-450LM2 molecule located close to the heme edge. AIA administration to the animals caused inhibition of hexobarbital-promoted electron flow from NADPH-cytochrome P-450 reductase to phenobarbital-inducible ferricytochrome P-450 both in microsomal particles and reconstituted systems. The impaired interaction between the proteins was shown not to originate from decreased capacity to bind each other but more likely to be due to some defect in a step subsequent to complex formation. In contrast, treatment with the porphyrogenic agent did not affect microsomal electron transmission from cytochrome b5 to the ferric monooxygenase. However, when the intermediate carrier was to donate reducing equivalents to the ferrous oxycytochrome in the presence of benzphetamine, there was a pronounced deceleration of the electron flux observable. These findings were interpreted to mean that there exist multiple reductase- and cytochrome-b5-binding domains in phenobarbital-inducible cytochrome P-450, some of which seem to be common to the two redox proteins. This sheds interesting light on the molecular organization of the catalytic electron transfer complexes.
Golly
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Walther Straub-Institut für Pharmakologie und Toxikologie, Universität, München, Germany.
Hlavica
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D000502:Allylisopropylacetamide / Q000494:pharmacology*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000737:chemistry; D015786:Cytochromes b5 / Q000737:chemistry; D004579:Electron Transport; D004790:Enzyme Induction; D006418:Heme / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010634:Phenobarbital; D011506:Proteins / Q000302:isolation & purification; D011817:Rabbits
D000596:Amino Acids; D065607:Cytochrome P-450 Enzyme Inhibitors; D011506:Proteins; D009243:NAD; D000502:Allylisopropylacetamide; D006418:Heme; D009249:NADP; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D010634:Phenobarbital