Comparison of some physicochemical and kinetic properties of S-adenosylhomocysteine hydrolase from bovine liver, bovine adrenal cortex and mouse liver.
S-Adenosyl-L-homocysteine hydrolase (EC 3.3.1.1) was purified to apparent homogeneity from bovine liver, bovine adrenal cortex and mouse liver. All enzymes were tetramers, composed of two types of subunit present in the proportion 1:1, as judged by SDS-polyacrylamide gel electrophoresis. The partition coefficient was exactly the same for these enzymes on high-performance gel permeation chromatography, and they co-sedimented in density gradients, suggesting the same molecular size and form of S-adenosylhomocysteine hydrolase from these sources. The bovine enzymes differed from the mouse liver enzyme with respect to isoelectric point (pI = 5.35, versus pI = 5.7), affinity for DEAE-cellulose, and migration of subunits on SDS-polyacrylamide gel electrophoresis with SDS from some commercial sources. The enzymes were not substrates for cAMP-dependent protein kinase. The apparent Km values for adenosine (0.2 microM) and S-adenosylhomocysteine (0.75 microM) were the same for all three enzymes. The ratio between Vmax for the synthesis and hydrolysis of S-adenosylhomocysteine was about 4 for the mouse liver enzyme, and about 6 for the bovine enzymes. It is concluded that only subtle kinetic and physicochemical differences exist between S-adenosylhomocysteine hydrolase from these bovine and mouse tissues. This suggests that differences in experimental procedures rather than species- and organ-differences of S-adenosylhomocysteine hydrolase are responsible for the variability in kinetic and physicochemical parameters reported for the mammalian hydrolase.
D043383:Adenosylhomocysteinase; D000302:Adrenal Cortex / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D007525:Isoelectric Focusing; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D051379:Mice; D011494:Protein Kinases / Q000378:metabolism; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D013045:Species Specificity
D012435:S-Adenosylhomocysteine; D000242:Cyclic AMP; D011494:Protein Kinases; D006867:Hydrolases; D043383:Adenosylhomocysteinase