Active Cytochrome P450 2E1 (CYP2E1)

Cytochrome P450 2E1 (CYP2E1) is a conserved microsomal monooxygenase predominantly expressed in the human liver, belonging to the cytochrome P450 superfamily. It mediates the oxidative metabolism of numerous small‐molecule xenobiotics, ethanol, and endogenous compounds, activating many procarcinogens and toxicants such as nitrosamines, benzene, and halogenated solvents. Besides metabolizing clinical drugs including acetaminophen and volatile anesthetics, CYP2E1 catalyzes redox reactions that generate reactive oxygen species (ROS), triggering oxidative stress, lipid peroxidation, and cellular damage. Its inducibility by alcohol and environmental chemicals links it to alcoholic liver injury, drug‐induced hepatotoxicity, and cancer susceptibility, making it a key enzyme in toxicology and disease research. Functionally, CYP2E1 activates toxic intermediates, while GSTM1 detoxifies these metabolites via glutathione conjugation.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human CYP2E1 and recombinant human GSTM1. Briefly, CYP2E1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GSTM1-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CYP2E1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant human CYP2E1 and recombinant human GSTM1 was shown in Figure 1, the EC50 for this effect is 0.076 µg/mL.