Uncoupling of cytochrome P450 1A and stimulation of reactive oxygen species production by co-planar polychlorinated biphenyl congeners.
The non-ortho-polychlorinated biphenyl (PCB) congener 3,3'4,4'-tetrachlorobiphenyl (PCB 77) can uncouple the catalytic cycle of fish (scup) cytochrome P4501A (CYP1A) and mammalian (rat, human) CYP1A1, stimulating release of reactive oxygen species (ROS). PCB 77 also inactivates CYP1A in an NADPH-, oxygen-, and time-dependent process, linked to uncoupling. We addressed a hypothesis that planar halogenated hydrocarbons generally will uncouple CYP1A. Thus, additional PCB congeners including non-ortho-3,3',4,4',5'-pentachlorobiphenyl (PCB 126) and 3,3',4,4',5,5'-hexachlorobiphenyl (PCB 169), mono-ortho-2,3,3',4,4'-pentachlorobiphenyl (PCB 105) and di-ortho-2,2',5,5'-tetrachlorobiphenyl (PCB 52), as well as the polycyclic aromatic hydrocarbon benzo[a]pyrene (B[a]P), were examined for their ability to stimulate microsomal ROS production and to inactivate CYP1A. Incubated without NADPH, non-ortho-PCB 126 and -PCB 169 both inhibited microsomal CYP1A activity (ethoxyresorufin O-deethylase; EROD). When NADPH was included, these congeners caused a progressive inactivation of CYP1A, in addition to the inhibition. The determined K(Inact) values for inactivation were 0.14 and 0.08 microM, respectively, for PCB 126 and PCB 169, similar to the 0.05 microM for PCB 77 previously reported. The mono-ortho-PCB 105 weakly inhibited and weakly inactivated CYP1A. The di-ortho-PCB 52 neither inhibited nor inactivated CYP1A. Alone, B[a]P strongly inhibited CYP1A, but when NADPH was added that inhibition was reversed, apparently by metabolic depletion of the substrate, and there was no inactivation. PCB 126 and PCB 169 stimulated release of ROS from induced liver microsomes, while B[a]P, PCB 52 and PCB 105 did not. ROS release and CYP1A inactivation stimulated by the non-ortho-PCB 126 and PCB 169 indicate an uncoupling of CYP1A like that previously shown with PCB 77. The uncoupling and release of ROS further suggest a participation of CYP1A in the oxidative stress associated with some planar halogenated aryl hydrocarbon receptor agonists.
Schlezinger
Jennifer J
JJ
Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
Struntz
William D J
WD
Goldstone
Jared V
JV
Stegeman
John J
JJ
D000818:Animals; D001386:Azides / Q000494:pharmacology; D001564:Benzo(a)pyrene / Q000494:pharmacology; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects* / Q000378:metabolism; D004579:Electron Transport / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D009249:NADP / Q000378:metabolism / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D010473:Perciformes / Q000378:metabolism*; D011078:Polychlorinated Biphenyls / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013053:Spectrophotometry / Q000662:veterinary; D014475:Uncoupling Agents / Q000494:pharmacology*
D001386:Azides; D065607:Cytochrome P-450 Enzyme Inhibitors; D017382:Reactive Oxygen Species; D014475:Uncoupling Agents; D001564:Benzo(a)pyrene; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D011078:Polychlorinated Biphenyls