Radioactive-electrophoretic assay of adenosine 5′-triphosphate sulfurylase activity in crude extracts with sulfate or selenate as a substrate
✍ Scribed by M.Leslie Hanna; Robert T. Taylor
- Book ID
- 102987407
- Publisher
- Elsevier Science
- Year
- 1989
- Tongue
- English
- Weight
- 968 KB
- Volume
- 176
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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.