Mechanism-based inactivation of human CYP2E1 by diethyldithocarbamate.
Although the ability of disulfiram to inactivate CYP2E1 has been known for more than 20 years, the mechanism has not yet been elucidated. A metabolite of disulfiram, diethyldithocarbamate (DDC), is converted by CYP2E1 to a reactive intermediate that subsequently inactivates the protein, leading to mechanism-based inactivation. Mass spectral analysis of the inactivated human 2E1 protein demonstrates that the inactivation is due to the formation of an adduct of the reactive metabolite of DDC with the apoprotein. These data, along with mass spectral analysis of a reactive intermediate trapped with GSH, indicate the involvement of a reactive intermediate with a molecular mass of 116 Da. Our results suggest that this binding involves formation of a disulfide bond with one of the eight cysteines in CYP2E1. The inactivation of wild-type CYP2E1 as well as two of its polymorphic mutants, CYP2E1*2 and CYP2E1*4, was also investigated. For wild-type CYP2E1, the K(I) was 12.2 μM and the k(inact) was 0.02 min(-1). The K(I) values for the two polymorphic mutants were 227.6 and 12.4 μM for CYP2E1.2 and CYP2E1.4, and the k(inact) values were 0.0061 and 0.0187 min(-1), respectively. These data indicate that DDC is a much less efficient inactivator of CYP2E1.2 than it is of either the wild-type or the CYP2E1.4 variant.
Pratt-Hyatt
Matthew
M
Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109-5632, USA.
Lin
Hsia-lien
HL
Hollenberg
Paul F
PF
D002851:Chromatography, High Pressure Liquid; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics; D065691:Cytochrome P-450 CYP2E1 Inhibitors; D004221:Disulfiram / Q000494:pharmacology; D004050:Ditiocarb / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D009154:Mutation; D053719:Tandem Mass Spectrometry
D065691:Cytochrome P-450 CYP2E1 Inhibitors; D004050:Ditiocarb; D019392:Cytochrome P-450 CYP2E1; D004221:Disulfiram
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