Liver microsomal epoxide hydrase.
1. The substrate specificity of membrane-bound and purified epoxide hydrase from rat liver microsomes has been studied. Both enzyme preparations catalyzed the hydration of a variety of alkene oxidase as well as arene oxides of several polycyclic aromatic hydrocarbons. 2. Unlike the membrane-bound enzyme, the rate of hydration for most of the substrates catalyzed by the purified epoxide hydrase was constant for only 1 or 2 min. The addition of dilauroyl phosphatidylcholine or heated microsomes to the incubation mixture extended the linearity of the reaction. 3. When rat liver microsomes were used as the source of the enzyme, the apparent Km values for many of the substrates were dependent on the amount of microsomes used. When purified epoxide hydrase was used as the enzyme source and benzo(a)pyrene 11,12-oxide as substrate, the apparent Km for benzo(a)pyrene 11,12-oxide was independent of enzyme concentration but dependent on added lipid concentration. Thus, in the absence of added dilauroyl phosphatidylcholine or in the presence of this lipid at a concentration below its critical micelle concentration, the observed Km for benzo(a)pyrene 11,12-oxide remained constant. However, when the lipid concentration was greater than the critical micelle concentration, the apparent Km value increased linearly with lipid concentration. These results are consistent with a model based on the partition of lipid-soluble substrate between the lipid micelle and the aqueous medium.
D000818:Animals; D004851:Epoxide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D006836:Hydro-Lyases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008566:Membranes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000494:pharmacology; D011083:Polycyclic Compounds / Q000378:metabolism*; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013329:Structure-Activity Relationship
D010713:Phosphatidylcholines; D011083:Polycyclic Compounds; D012710:Serum Albumin, Bovine; D004851:Epoxide Hydrolases; D006836:Hydro-Lyases