Distribution and nature of epoxide hydrolase activity in subcellular organelles of mouse liver.
Mouse liver light and heavy mitochondrial fractions contain significant epoxide hydrolase activity in addition to that present in the cytosol and microsomes. As the mitochondrial fraction itself contains a number of subfractions, experiments were designed to determine the localization of the epoxide hydrolase activity in these subfractions. Subcellular fractions were prepared using livers from 6- to 8-week-old Swiss-Webster male mice. Using trans-stilbene oxide (TSO) as substrate, the highest activity was localized in the cytosolic fraction, followed by the light mitochondrial fraction. Subfractionation of the light mitochondrial fraction by isopycnic sucrose density gradient resulted in the separation of mitochondria from peroxisomes as monitored by marker enzymes. The separation of these two subcellular organelles was also confirmed by the electron microscopic studies. Distribution of TSO-hydrolase activity in the sucrose density gradient fractions closely resembled the activity distribution of the peroxisomal markers catalase and urate oxidase, but significant activity was also found in mitochondria. Treatment of mice with clofibrate selectively induced TSO-hydrolase in the cytosol without affecting this enzyme activity in the peroxisomal fraction. There was no difference in the distribution pattern of TSO-hydrolase and marker enzymes in sucrose density gradients of mitochondrial fractions from clofibrate-treated and control mice. The epoxide hydrolase activity in the peroxisomes is immunologically similar to, and also has the same molecular weight as, the cytosolic epoxide hydrolase.
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D000818:Animals; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D002994:Clofibrate / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D004852:Epoxy Compounds / Q000378:metabolism; D005779:Immunodiffusion; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D051379:Mice; D008830:Microbodies / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D008995:Monoamine Oxidase / Q000032:analysis; D011506:Proteins / Q000032:analysis; D011817:Rabbits; D013267:Stilbenes / Q000378:metabolism; D013385:Succinate Dehydrogenase / Q000032:analysis
D004852:Epoxy Compounds; D011506:Proteins; D013267:Stilbenes; D013385:Succinate Dehydrogenase; D008995:Monoamine Oxidase; D004851:Epoxide Hydrolases; D002994:Clofibrate; C025906:stilbene oxide