一种用于检测硒磷酸合成酶活性的非放射性和两种放射性检测方法。
A non-radioactive and two radioactive assays for selenophosphate synthetase activity.
作者信息
Liu S Y, Stadtman T C
机构信息
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
出版信息
Biofactors. 1997;6(3):305-9. doi: 10.1002/biof.5520060301.
Selenophosphate synthetase catalyzes the formation of monoselenophosphate (SePO3(3-)) from ATP and selenide (reaction 1). [formula: see text] In one assay frequently used, [8-14C]AMP formation from [8-14C]ATP is estimated after separation of the nucleotides by thinlayer chromatography. An alternative non-radioactive assay in which the AMP product is estimated using AMP deaminase is described. The highly oxygen-labile selenophosphate product can be estimated in an assay employing [gamma-32P]ATP. The 32P-labeled selenophosphate is converted to [32P]orthophosphate by treatment with iodine and estimated after removal of residual [32P]ATP on charcoal.
硒磷酸合成酶催化由ATP和硒化物形成单硒磷酸酯（SePO3(3-)）（反应1）。[化学式：见正文]在一种常用的测定方法中，通过薄层层析分离核苷酸后，估计[8-14C]ATP形成[8-14C]AMP的情况。本文描述了另一种使用AMP脱氨酶估计AMP产物的非放射性测定方法。在使用[γ-32P]ATP的测定中，可以估计对氧高度不稳定的硒磷酸酯产物。用碘处理后，32P标记的硒磷酸酯转化为[32P]正磷酸盐，并在通过活性炭去除残留的[32P]ATP后进行估计。
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