The effect of detergents on the purified flavin-containing monooxygenase of mouse liver, kidney and lungs.
1. The effect of various commonly used membrane solubilizing detergents on the activity of the microsomal xenobiotic metabolizing enzyme, the flavin-containing monooxygenase (FMO) purified from mouse liver, kidney and lungs was determined. 2. Regardless of the type of detergent used, the effect on the enzyme activity was variable depending on the type of substrate used. 3. Emulgen 911 concentrations of up to 10% had very little effect on thiobenzamide-S-oxidation by liver, kidney or lung FMO. 4. While Emulgen 911 increased substrate dependent NADPH oxidation rate by thiourea and thioacetamide, it drastically reduced the activity toward the organophosphorous compounds, disulfoton, fenthion, fonofos and phorate at low concentrations. 5. Activities of fenthion, phorate and fonofos were decreased by 80, 65 and 55% by the inclusion of 0.25% Emulgen 911 in the assay mixture. 6. This decline in FMO activity for phorate was evident regardless of the type of detergent used. In contrast, thiourea dependent NADPH oxidation rate in the presence of various detergents was variable. 7. Thiourea oxidation rate was decreased by cholate and Zwittergent 3-12, whereas it was increased in the presence of Emulgen 911, Triton X-100 and Tween 20. 8. This study shows that before FMO activity is determined in the presence of detergents their effects should be carefully evaluated.
Venkatesh
K
K
Department of Toxicology, North Carolina State University, Raleigh 27695.
Levi
P E
PE
Hodgson
E
E
D000818:Animals; D065607:Cytochrome P-450 Enzyme Inhibitors; D003902:Detergents / Q000494:pharmacology*; D066298:In Vitro Techniques; D007668:Kidney / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008168:Lung / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D051379:Mice; D009249:NADP / Q000378:metabolism; D017137:Nonoxynol; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000037:antagonists & inhibitors*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D013890:Thiourea / Q000378:metabolism
D065607:Cytochrome P-450 Enzyme Inhibitors; D003902:Detergents; D017137:Nonoxynol; D011092:Polyethylene Glycols; D009249:NADP; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); D013890:Thiourea