DEVELOPMENT OF NIR FOR MONITORING ENZYME-SUBSTRATE REACTIONS

用Na-K ATP酶测定了ATP水解的动力学。 NIR光谱法。 先前的结果已经确定， 平衡溶液中ATP、ADP和PO 4的量可以是 用NIR光谱定量测定。 的主要优点 使用NIR光谱是该方法的时间分辨能力， 使得光谱学家能够在非常快的时间内监测反应 鳞片 考虑到这一点，我们已经观察到ATP的水解， 在十秒时间范围内的近红外光谱。 变化 近红外光谱区域反映了ATP酶水解为ADP。 将这些光谱变化分离为它们的单独分量， 已经完成;然而，光密度的整体变化 与无机磷酸盐的独立测定线性相关。 与 随着近红外技术的迅速发展， 时间分辨率的扩展以及 组件有望实现。

The kinetics of the ATP hydrolysis by Na-K ATPase has been monitored with NIR spectroscopy. Previous results have determined that the relative amounts of ATP, ADP, and PO4 in equilibria solutions can be quantitatively determined with NIR spectroscopy. The major advantage of using NIR spectroscopy is the time-resolved capability of the method, allowing the spectroscopist to monitor reactions at very fast time scales. With this in mind, we have observed the hydrolysis of ATP with near infrared spectroscopy in the ten second time regime. Changes in the NIR spectral region reflect the enzymatic hydrolysis of ATP to ADP. Separation of these spectral changes to their separate components has not been accomplished; however, the overall change of optical density correlates linearly with independent assay of inorganic phosphate. With the advances in near infrared technology rapidly progressing, the extension of the temporal resolution as well as the separation of components is hoped to be achieved.