Assay and regulation of S-adenosylmethionine synthetase in Saccharomyces cerevisiae and Candida utilis.
A simple and sensitive assay for S-adenosylmethionine (SAM) synthetase is described which depends on the quantitative separation of the product, [14CH3]S-adenosylmethionine, from the substrate, L-[14CH3]methionine, on a Bio-Rex 70 column. L-Methionine protects the enzyme during preparation of cell extracts by sonic treatment but causes repression of enzyme activity during growth of Candida utilis. The presence of 5 mM methionine in the growth medium repressed SAM synthetase specific activity threefold compared to the specific acitivity of the enzyme isolated from cells grown in unsupplemented medium. Conversely, the presence of methionine in the growth medium resulted in an 80-fold increase in the intracellular concentration of SAM as compared to the Sam accumulated intracellularly in unsupplemented cultures.
D000255:Adenosine Triphosphate; D002175:Candida / Q000201:enzymology* / Q000378:metabolism; D002250:Carbon Radioisotopes; D002474:Cell-Free System; D002852:Chromatography, Ion Exchange; D004794:Enzyme Repression; D008715:Methionine / Q000378:metabolism; D010710:Phosphates / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D014166:Transferases / Q000378:metabolism*
D002250:Carbon Radioisotopes; D010710:Phosphates; D013912:Threonine; D012436:S-Adenosylmethionine; D000255:Adenosine Triphosphate; D008715:Methionine; D014166:Transferases