Partial purification and characterization of S-adenosylhomocysteine hydrolase isolated from Saccharomyces cerevisiae.
S-Adenosylhomocysteine (SAH) hydrolase was purified 25-fold from bakers' yeast by chemical methods and column chromatography. The purified enzyme could readily synthesize SAH from adenosine and homocysteine, but could hydrolyze only negligible amounts of SAH. The purified enzyme showed no activity towards S-adenosylmethionine, methylthioadenosine, or adenosine. Several nucleotides, sulfhydryl compounds, and ribose could not replace adenosine or homocysteine in the reaction mixture. SAH could be hydrolyzed by SAH hydrolase if commercial adenosine deaminase was included in the reaction mixture. Under these conditions l-homocysteine could act as a product inhibitor. A number of compounds structurally similar to adenosine and homocysteine were found to inhibit synthesis of SAH from adenosine and homocysteine. The strongest inhibitors were adenine, adenosine-3'-monophosphate, adenosine-2'-monophosphate, adenosine diphosphate, adenosine triphosphate, and adenosine-5'-monophosphate. The biosynthetic and hydrolytic activity of SAH hydrolase in yeast cell ghosts was similar to the activity of the enzyme in vitro.
D000096:Acetone; D000225:Adenine / Q000494:pharmacology; D000241:Adenosine / Q000378:metabolism; D000619:Aminohydrolases / Q000378:metabolism; D002247:Carbon Isotopes; D002474:Cell-Free System; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002854:Chromatography, Paper; D003911:Dextrans; D006710:Homocysteine / Q000378:metabolism; D006867:Hydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D006882:Hydroxyapatites; D009705:Nucleosides / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D012263:Ribonucleosides / Q000096:biosynthesis / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014461:Ultracentrifugation
D002247:Carbon Isotopes; D003911:Dextrans; D006882:Hydroxyapatites; D009705:Nucleosides; D009711:Nucleotides; D012263:Ribonucleosides; D006710:Homocysteine; D000096:Acetone; D006867:Hydrolases; D000619:Aminohydrolases; D000225:Adenine; D000241:Adenosine