Multiple species of mammalian S-adenosylmethionine synthetase. Partial purification and characterization.
Two species of S-adenosylmethionine (S-Ado-Met) synthetase (EC 2.5.1.6) exist in rat liver cytosol and a distinct species of the enzyme exists in kidney cytosol. S-Ado-Met synthetases alpha and beta in rat liver cytosol have been partially purified about 200- and 80-fold, respectively. The apparent molecular weight estimated by gel filtration and the sedimentation coefficient are 210 000 and 9 S for S-Ado-Met synthetase alpha and 160 000 and 5.5 S for S-Ado Met synthetase beta. Both enzymes absolutely require Mg2+ and K+ for the activity and are completely inhibited by p-(chloromercuri)-benzoate. Kinetic studies indicate that S-Ado-Met synthetases alpha and beta exhibit negative cooperativity with low S0.5 (ligand concentration required for half-maximal velocity) for L-methionine (17 microM) and ATP (0.5 mM) and positive cooperativity with much higher S0.5 values (S0.5 (L-methionine) = 0.5 mM, S0.5 (ATP) = 2 mM), respectively. The cryoprotectants dimethyl sulfoxide and glycerol markedly lower the S0.5 values of S-Ado-Met synthetase beta without significant effect on Vmax. A single species of S-Ado-Met synthetase has been purified about 1000-fold from rat kidney cytosol. The kidney enzyme, termed S-Ado-Met synthetase gamma, has an apparent molecular weight of 190 000 and a sedimentation coefficient of 7.5 S and is resistant to the inhibition by p-(chloromercuri)benzoate. S-Ado-Met synthetase gamma exhibits slightly negative cooperativity with an apparent S0.5 value for L-methionine of 6 microM and for ATP of 70 microM.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000302:isolation & purification*; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008274:Magnesium; D008715:Methionine / Q000378:metabolism; D008716:Methionine Adenosyltransferase / Q000302:isolation & purification*; D008970:Molecular Weight; D011188:Potassium; D051381:Rats; D014018:Tissue Distribution; D014166:Transferases / Q000302:isolation & purification*
D007527:Isoenzymes; D000255:Adenosine Triphosphate; D008715:Methionine; D014166:Transferases; D008716:Methionine Adenosyltransferase; D008274:Magnesium; D011188:Potassium