S-adenosylmethionine decarboxylase from baker's yeast.
1. S-Adenosyl-L-methionine decarboxylase (S-adenosyl-L-methionine carboxy-lyase, EC 4.1.1.50) was purified more than 1100-fold from extracts of Saccharomyces cerevisiae by affinity chromatography on columns of Sepharose containing covalently bound methylglyoxal bis(guanylhydrazone) (1,1'[(methylethanediylidene)dinitrilo]diguanidine) [Pegg, (1974) Biochem J. 141, 581-583]. The final preparation appeared to be homogeneous on polyacrylamide-gel electrophoresis at pH 8.4. 2. S-Adenosylmethionine decarboxylase activity was completely separated from spermidine synthase activity [5'-deoxyadenosyl-(5'),3-aminopropyl-(1),methylsulphonium-salt-putrescine 3-aminopropyltransferase, EC 2.5.1.16] during the purification procedure. 3. Adenosylmethionine decarboxylase activity from crude extracts of baker's yeast was stimulated by putrescine, 1,3-diamino-propane, cadaverine (1,5-diaminopentane) and spermidine; however, the purified enzyme, although still stimulated by the diamines, was completely insensitive to spermidine. 4. Adenosylmethionine decarboxylase has an apparent Km value of 0.09 mM for adenosylmethionine in the presence of saturating concentrations of putrescine. The omission of putrescine resulted in a five-fold increase in the apparent Km value for adenosylmethionine. 5. The apparent Ka value for putrescine, as the activator of the reaction, was 0.012 mM. 6. Methylglyoxal bis(guanylhydrazone) and S-methyladenosylhomocysteamine (decarboxylated adenosylmethionine) were powerful inhibitors of the enzyme. 7. Adenosylmethionine decarboxylase from baker's yeast was inhibited by a number of conventional carbonyl reagents, but in no case could the inhibition be reversed with exogenous pyridoxal 5'-phosphate.
D012437:Adenosylmethionine Decarboxylase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D001596:Benzylamines / Q000494:pharmacology; D001894:Borohydrides / Q000037:antagonists & inhibitors; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003486:Cyanides / Q000494:pharmacology; D003959:Diamines / Q000494:pharmacology; D007538:Isoniazid / Q000494:pharmacology; D008935:Mitoguazone; D011700:Putrescine / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012663:Semicarbazides / Q000494:pharmacology; D012685:Sepharose; D013095:Spermidine / Q000494:pharmacology; D011436:Spermidine Synthase / Q000378:metabolism
D001596:Benzylamines; D001894:Borohydrides; D003486:Cyanides; D003959:Diamines; D012663:Semicarbazides; D011732:Pyridoxal Phosphate; D012685:Sepharose; D011436:Spermidine Synthase; D002262:Carboxy-Lyases; D012437:Adenosylmethionine Decarboxylase; D008935:Mitoguazone; D013095:Spermidine; D011700:Putrescine; D007538:Isoniazid