Adrenal activation of carbon tetrachloride: role of microsomal P450 isozymes.
Previous investigations demonstrated that carbon tetrachloride (CCl4) was activated by adrenal microsomes, resulting in various functional changes and ultimately in necrosis of the zona reticularis of the gland. Experiments were done to identify the adrenal P450 isozyme(s) involved in the bioactivation of CCl4. Incubation of microsomes from the zona reticularis (ZR) of the guinea pig adrenal cortex with CCl4 plus NADPH caused initiation of lipid peroxidation, covalent binding of CCl4-derived radioactivity to protein, and degradation of cytochrome(s) P450. Preincubation of the microsomal preparations with inhibitory antibodies to P450(17 alpha) or P450C21 decreased the corresponding enzyme activities (17 alpha-hydroxylation and 21-hydroxylation), but did not affect the activation of CCl4. 1-Aminobenzotriazole (ABT), a suicide inhibitor of some P450 isozymes, decreased the enzyme activities catalysed by an adrenal 52,000 Da (52 kDa) isozyme, but had no effect on the function of P450(17 alpha) or P450C21. However, ABT completely inhibited the CCl4-induced LP and covalent binding in adrenal microsomes. The results indicate that adrenal CCl4 activation is catalysed by the 52 kDa P450 isozyme and not by the steroid hydroxylases. Localization of the 52 kDa isozyme to the ZR probably accounts for the selective necrosis of this region of the gland by CCl4.
Colby
H D
HD
Department of Pharmacology and Toxicology, Philadelphia College of Pharmacy and Science, PA 19104.
Purcell
H
H
Kominami
S
S
Takemori
S
S
Kossor
D C
DC
D000302:Adrenal Cortex / Q000187:drug effects* / Q000201:enzymology*; D000818:Animals; D002251:Carbon Tetrachloride / Q000378:metabolism / Q000633:toxicity*; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006168:Guinea Pigs; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D008861:Microsomes / Q000187:drug effects* / Q000378:metabolism; D009249:NADP / Q000494:pharmacology; D014230:Triazoles / Q000494:pharmacology
D065607:Cytochrome P-450 Enzyme Inhibitors; D007527:Isoenzymes; D014230:Triazoles; C033020:1-aminobenzotriazole; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D002251:Carbon Tetrachloride