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 ATPase的ATP水解动力学已通过 NIR光谱。 先前的结果已经确定了相对 平衡解决方案中的ATP，ADP和PO4量可以是 用NIR光谱法定量确定。 主要优势 使用NIR光谱是该方法的时间分辨能力， 允许光谱学家在很快的时间监测反应 秤。 考虑到这一点，我们已经观察到ATP的水解 在十次第二次制度中，近红外光谱法。 变化 NIR光谱区域反映了ATP至ADP的酶促水解。 将这些光谱变化与它们的单独组件的分离尚未 已完成；但是，光密度的总体变化 与无机磷酸盐的独立测定线性相关。 和 近红外技术的进步迅速发展， 时间分辨率的扩展以及分离 希望要实现组件。