Radioactive-electrophoretic assay of adenosine 5'-triphosphate sulfurylase activity in crude extracts with sulfate or selenate as a substrate.
An assay method for ATP sulfurylase is presented which employs Na2(35)SO4 as a substrate and measures the production of labeled adenosine 5'-phosphosulfate and 3'-phosphoadenosine 5'-phosphosulfate by low-voltage, hanging paper strip electrophoresis. The method is applicable to crude bacterial or mammalian extracts and accurately measures picomole amounts of product(s). Na2(75SeO4 can also be employed as a substrate, if the unstable radioactive product, adenosine 5'-phosphoselenate, is converted to elemental 75Se degrees by inclusion of reduced glutathione in the reaction mixture. The same paper strip electrophoretic technique can then be used to separate 75Se degrees from the radiolabeled substrate. The method also has utility for measuring any direct reduction by crude microbial extracts of radioactive selenate to selenite, independent of ATP sulfurylase.
Hanna
M L
ML
Biomedical Sciences Division, Lawrence Livermore National Laboratory, University of California, Livermore 94550.
Taylor
R T
RT
D000250:Adenosine Phosphosulfate / Q000032:analysis; D000818:Animals; D003013:Clostridium / Q000201:enzymology; D006224:Cricetinae; D004926:Escherichia coli / Q000201:enzymology; D005233:Fatty Alcohols / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000032:analysis*; D010724:Phosphoadenosine Phosphosulfate / Q000032:analysis; D012643:Selenium / Q000378:metabolism*; D018036:Selenium Compounds; D064588:Selenium Oxides; D012644:Selenium Radioisotopes; D012981:Sodium Tetradecyl Sulfate / Q000378:metabolism*; D013430:Sulfate Adenylyltransferase / Q000032:analysis*; D013460:Sulfur Isotopes; D013855:Thiobacillus / Q000201:enzymology
D005233:Fatty Alcohols; D018036:Selenium Compounds; D064588:Selenium Oxides; D012644:Selenium Radioisotopes; D013460:Sulfur Isotopes; D010724:Phosphoadenosine Phosphosulfate; D000250:Adenosine Phosphosulfate; D009713:Nucleotidyltransferases; D013430:Sulfate Adenylyltransferase; D012643:Selenium; D012981:Sodium Tetradecyl Sulfate