Polyamine synthesis in mammalian tissues. Isolation and characterization of spermidine synthase from bovine brain.
Spermidine synthase (EC 2.5.1.16) was purified to apparent homogeneity (about 11 000-fold) from bovine brain by affinity chromatography, with S-adenosyl-(5')-3-thiopropylamine linked to Sepharose as the adsorbent. The enzyme preparation was free from S-adenosylmethionine decarboxylase (EC 4.1.1.50) and spermine synthase (EC 2.5.1.22) activities. The native enzyme had an apparent Mr of 70 000, was composed of two subunits of equal size, and had an isoelectric point at pH 5.22. The apparent Km values for putrescine and decarboxylated adenosylmethionine [S-adenosyl-(5')-3-methylthiopropylamine] were 40 microM and 0.3 microM respectively. Cadaverine and 1,6-diaminohexane could replace putrescine as the aminopropyl acceptor, although the reaction rates were only 6% and 1% respectively of that obtained with putrescine. Ethyl, propyl and carboxymethyl analogues of decarboxy-S-adenosylmethionine could act as propylamine donors. Both the reaction products, spermidine and 5'-methylthioadenosine, were mixed-type inhibitors of the enzyme. On the basis of initial-velocity and product-inhibition studies, a ping-pong reaction mechanism for the spermidine synthase reaction was ruled out.
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D003839:Deoxyadenosines; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D011700:Putrescine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D013095:Spermidine / Q000494:pharmacology; D011436:Spermidine Synthase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D013872:Thionucleosides / Q000494:pharmacology; D014166:Transferases / Q000302:isolation & purification*
D003839:Deoxyadenosines; D013872:Thionucleosides; C008500:5'-methylthioadenosine; D012436:S-Adenosylmethionine; D014166:Transferases; D011436:Spermidine Synthase; D000241:Adenosine; D013095:Spermidine; D011700:Putrescine