title: Spectrophotometric analysis of human CYP2E1-catalyzed p-nitrophenol hydroxylation.
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.
authors.affiliation: Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, Canada.
authors.forename: Thomas K H
authors.initials: TK
authors.lastname: Chang
authors.forename: Charles L
authors.initials: CL
authors.lastname: Crespi
authors.forename: David J
authors.initials: DJ
authors.lastname: Waxman
mesh_terms: D002384:Catalysis; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D008862:Microsomes, Liver / Q000201:enzymology; D009596:Nitrophenols / Q000378:metabolism*
chemical_list: D009596:Nitrophenols; D019392:Cytochrome P-450 CYP2E1; C024836:4-nitrophenol