Pyrophosphate and tripolyphosphate affect firefly luciferase luminescence because they act as substrates and not as allosteric effectors
Pyrophosphate and tripolyphosphate affect firefly luciferase luminescence because they act as substrates and not as allosteric effectors
复制标题
DOI：
10.1111/j.1742-4658.2008.06309.x
复制
发表时间：
2008-04-01
期刊：
影响因子：
5.4
通讯作者：
da Silva, Joaquim C. G. Esteves
中科院分区：
文献类型：
--
作者：
Fontes, Rui;Fernandes, Diogo;da Silva, Joaquim C. G. Esteves
研究了无机焦磷酸盐、三聚磷酸盐和核苷三磷酸盐对萤火虫荧光素酶生物发光的激活和稳定作用。所获得的结果表明，这些效果是一个后果的脱氢酶催化分裂的脱氢腺苷酸，一个强大的抑制剂形成的生物发光反应过程中的副产物。无机焦磷酸盐、三聚磷酸盐、CTP和UTP拮抗脱氢乙酰基-腺苷酸的抑制作用，因为它们与其反应产生至少是较弱抑制剂的产物。此外，我们表明，拮抗作用取决于分裂反应的速率较高的无机焦磷酸盐和三聚磷酸盐的情况下，CTP和UTP的情况下，较低。在无机焦磷酸盐的情况下，之间的相关性的脱氢乙酰基-腺苷酸焦磷酸解的速率和对生物发光的激活作用只发生在低浓度，因为无机焦磷酸盐是，同时，生物发光反应的抑制剂。我们的研究结果表明，以前的报告有关的几种核苷酸（包括一些不与脱氢乙酰腺苷酸反应）对生物发光的激活作用所造成的无机焦磷酸盐污染的存在下使用的制剂。
The activating and stabilizing effects of inorganic pyrophosphate, tripolyphosphate and nucleoside triphosphates on firefly luciferase bioluminescence were studied. The results obtained show that those effects are a consequence of the luciferase-catalyzed splitting of dehydroluciferyl-adenylate, a powerful inhibitor formed as a side product in the course of the bioluminescence reaction. Inorganic pyrophosphate, tripolyphosphate, CTP and UTP antagonize the inhibitory effect of dehydroluciferyl-adenylate because they react with it giving rise to products that are, at least, less powerful inhibitors. Moreover, we demonstrate that the antagonizing effects depended on the rate of the splitting reactions being higher in the cases of inorganic pyrophosphate and tripolyphosphate and lower in the cases of CTP and UTP. In the case of inorganic pyrophosphate, the correlation between the rate of dehydroluciferyl-adenylate pyrophosphorolysis and the activating effect on bioluminescence only occurs for low concentrations because inorganic pyrophosphate is, simultaneously, an inhibitor of the bioluminescence reaction. Our results demonstrate that previous reports concerning the activating effects of several nucleotides (including some that do not react with dehydroluciferyl-adenylate) on bioluminescence were caused by the presence of inorganic pyrophosphate contamination in the preparations used.