Regulation of S-adenosylmethionine decarboxylase activity in rat liver and prostate.
Two methods were used for the quantitation of S-adenosylmethionine decarboxylase protein. The first involved titrating the active site of the enzyme by reduction of the Schiff base between 3H-decarboxylated S-adenosylmethionine and the pyruvate prosthetic group with sodium cyanoborohydride. The second method was radioimmunoassay with rabbit antiserum which was used to determine the total immunoreactive enzyme protein. It was found that the increased S-adenosylmethionine decarboxylase activity produced in rat prostate by treatment with alpha-difluoromethylornithine and in both prostate and liver by methylglyoxal bis(guanylhydrazone) were due entirely to increases in the amount of enzyme protein. The ratio of enzyme activity to protein (measured by either method) remained constant in rats treated with the drugs. Treatment with 2% alpha-difluoromethylornithine in the drinking water for 3 days increased prostatic S-adenosylmethionine decarboxylase protein by 5-fold. A substantial part, but not all, of this increase could be accounted for by a slowing of the rate of degradation of the enzyme. The half-life for loss of activity and titratable protein after inhibition of protein synthesis by cycloheximide was increased from 35 to 108 min by treatment with alpha-difluoromethylornithine. However, the half-life for loss of immunoreactive protein which was considerably longer was only increased from 139 to 213 min. The molecular weight of the S-adenosylmethionine decarboxylase subunit determined by immunoblotting was 32,000, and no smaller immunoreactive fragments were detected. These results indicate that spermidine depletion produced by alpha-difluoromethylornithine affects the degradation of S-adenosylmethionine decarboxylase at an early step involving the loss of the active site without substantial breakdown of the protein.
D012437:Adenosylmethionine Decarboxylase / Q000378:metabolism*; D000818:Animals; D002262:Carboxy-Lyases / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D000518:Eflornithine; D006207:Half-Life; D007163:Immunosorbent Techniques; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008935:Mitoguazone / Q000494:pharmacology; D008970:Molecular Weight; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D011467:Prostate / Q000187:drug effects / Q000201:enzymology*; D011700:Putrescine / Q000378:metabolism; D011817:Rabbits; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013095:Spermidine / Q000378:metabolism; D013096:Spermine / Q000378:metabolism
D013096:Spermine; D003513:Cycloheximide; D009952:Ornithine; D002262:Carboxy-Lyases; D012437:Adenosylmethionine Decarboxylase; D008935:Mitoguazone; D013095:Spermidine; D011700:Putrescine; D000518:Eflornithine