Biosynthesis of enzymes of rat-liver mitochondrial beta-oxidation.
The biogenesis of seven enzymes involved in the mitochondrial fatty acid beta-oxidation of rat liver was studied. Hepatic RNA was translated in vitro in a rabbit reticulocyte lysate cell-free system and the translation products were immunoprecipitated, subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and visualized by fluorography. The translation products obtained in vitro of medium-chain and/or long-chain acyl-CoA dehydrogenase (these enzymes were immunochemically cross-reactive), enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and acetoacetyl-CoA thiolase and probably also short-chain acyl-CoA dehydrogenase were larger than the subunits of the corresponding mature enzymes by 2-4.5 kDa, whereas the 3-oxoacyl-CoA thiolase obtained in vitro was approximately the same size as the mature subunit. The free polysome fraction of rat liver was 4.3-9.0-times more active than the membrane-bound polysome fraction in the synthesis of these seven enzymes. The enzyme activities were increased after administration of di(2-ethylhexyl)phthalate; the extent of the increase varied from one enzyme to another. The increase in the cell-free translation activity of total hepatic RNA for these enzymes after administration of the chemical was markedly different among individual enzymes and higher than that in the rates of synthesis of the corresponding enzymes which were determined by the experiment in vivo.
D000818:Animals; D002474:Cell-Free System; D004051:Diethylhexyl Phthalate / Q000494:pharmacology; D004790:Enzyme Induction; D005227:Fatty Acids / Q000378:metabolism*; D005470:Fluorometry; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D005227:Fatty Acids; D012333:RNA, Messenger; D004051:Diethylhexyl Phthalate