Abstract
The cytochrome P450 enzyme CYP2E1 catalyzes the oxidative metabolism of many solvents and other small organic molecules. A spectrophotometric method is described for determination of CYP2E1 activity by monitoring the formation of p-nitrocatechol from p-nitrophenol by cDNA-expressed CYP2E1 or isolated liver microsomes. The enzymatic product, p-nitrocatechol, is assayed at 535 nm after acidification of the reaction mixture with trichloroacetic acid followed by neutralization using 2 M NaOH. This method is applicable to enzymatic studies for determination of P450-catalyzed p-nitrophenol hydroxylation activity.
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Acknowledgments
This work was supported in part by the Canadian Institutes of Health Research (grant MOP-42385) and the National Institutes of Health (grant 5 P42 ES07381).
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Chang, T.K.H., Crespi, C.L., Waxman, D.J. (2006). Spectrophotometric Analysis of Human CYP2E1-Catalyzed p-Nitrophenol Hydroxylation. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology, vol 320. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-998-2:127
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DOI: https://doi.org/10.1385/1-59259-998-2:127
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