Investigation of structure and rate of synthesis of ornithine decarboxylase protein in mouse kidney.
An immunoblotting technique was used to study the forms of ornithine decarboxylase present in androgen-induced mouse kidney. Two forms were detected which differed slightly in isoelectric point but not in subunit molecular weight (approximately 55 000). Both forms were enzymatically active and could be labeled by reaction with radioactive alpha-(difluoromethyl)-ornithine, an enzyme-activated irreversible inhibitor. On storage of crude kidney homogenates or partially purified preparations of ornithine decarboxylase, the enzyme protein was degraded to a smaller size (Mr approximately 53 000) without substantial loss of enzyme activity. The synthesis and degradation of ornithine decarboxylase protein were studied by labeling the protein by intraperitoneal injection of [35S]methionine and immunoprecipitation using both monoclonal and polyclonal antibodies. The fraction of total protein synthesis represented by renal ornithine decarboxylase was increased at least 25-fold by testosterone treatment of female mice and was found to be about 1.1% in the fully induced androgen-treated female. Both forms of the enzyme were rapidly labeled in vivo, and the immunoprecipitable ornithine decarboxylase protein was almost completely lost after 4-h exposure to cycloheximide, confirming directly the very rapid turnover of this enzyme. Treatment with 1,3-diaminopropane which is known to cause a great reduction in ornithine decarboxylase activity did not greatly selectively inhibit the synthesis of the enzyme. However, 1,3-diaminopropane did produce an increase in the rate of degradation of ornithine decarboxylase and a general reduction in protein synthesis. These two factors, therefore, appear to be responsible for the loss of ornithine decarboxylase activity and protein in response to 1,3-diaminopropane.
D000818:Animals; D000911:Antibodies, Monoclonal; D003959:Diamines / Q000494:pharmacology; D004790:Enzyme Induction; D005260:Female; D007527:Isoenzymes / Q000302:isolation & purification; D007668:Kidney / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009955:Ornithine Decarboxylase / Q000096:biosynthesis*; D011863:Radioimmunoassay; D013462:Sulfur Radioisotopes; D013739:Testosterone / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D003959:Diamines; D007527:Isoenzymes; D013462:Sulfur Radioisotopes; D013739:Testosterone; C009475:trimethylenediamine; D009955:Ornithine Decarboxylase