Kinetics of a self-amplifying substrate cycle:: ADP-ATP cycling assay
Kinetics of a self-amplifying substrate cycle:: ADP-ATP cycling assay
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DOI：
10.1042/0264-6021:3500237
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发表时间：
2000-08-15
影响因子：
4.1
通讯作者：
García-Carmona, F
中科院分区：
文献类型：
--
作者：
Valero, E;Varón, R;García-Carmona, F
本文研究了腺苷酸激酶、丙酮酸激酶和L-乳酸脱氢酶参与的ATP-ADP扩增循环系统的动力学。循环的化学计量比为2：1，因为两个ADP分子由ATP和AMP中的每一个合成，并且在循环的每一个回合中，一个ADP分子被转化回一个ATP。根据获得的方程，这导致反应介质中ATP和ADP的浓度连续指数增加。因此，这是一个放大自身的基质循环，循环速率随时间连续增加。通过使用腺苷三磷酸双磷酸酶降解试剂中的ATP和ADP来降低试剂的背景信号，允许在连续分光光度测定中检测限低至16 pmol的ATP和/或ADP。
A kinetic study of an ATP-ADP amplification cyclic system involving the enzymes adenylate kinase, pyruvate kinase and L-lactate dehydrogenase has been made. The stoichiometry of the cycle is 2:1, because two molecules of ADP are synthesized from one each of ATP and AMP, and one molecule of ADP is converted back into one of ATP at each turn of the cycle. This results in a continuous exponential increase in the concentrations of ATP and ADP in the reaction medium, according to the equations obtained. This is therefore a substrate cycle that amplifies itself, the cycling rate increasing continuously with time. The background signal of the reagent was reduced by using apyrase to degrade ATP and ADP in the reagent, permitting detection limits as low as 16 pmol of ATP and/or ADP in a continuous spectrophotometric assay.