DEVELOPMENT OF NIR FOR MONITORING ENZYME-SUBSTRATE REACTIONS

开发用于监测酶-底物反应的近红外光谱

• 批准号: 3757623
• 负责人: R L BERGER
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3757623
• 关键词: adenosine diphosphate; adenosine triphosphate; bioengineering /biomedical engineering; biomedical equipment development; chemical kinetics; enzyme activity; enzyme substrate complex; hydrolysis; infrared spectrometry; phosphorus

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.

Na-K ATP 酶水解 ATP 的动力学已通过以下方法监测 近红外光谱。 先前的结果已确定相对 平衡溶液中 ATP、ADP 和 PO4 的量可 采用近红外光谱法进行定量测定。 主要优点 使用近红外光谱法是该方法的时间分辨能力， 允许光谱学家非常快地监测反应 秤。 考虑到这一点，我们观察了 ATP 的水解 十秒时间范围内的近红外光谱。 的变化 NIR 光谱区域反映了 ATP 酶促水解为 ADP。 尚未将这些光谱变化分离到其单独的成分中 已完成；然而，光密度的总体变化 与无机磷酸盐的独立测定呈线性相关。 和 随着近红外技术的飞速发展， 时间分辨率的扩展以及分离 组件有望实现。