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.

测定了Na-K-ATPase催化ATP水解酶的动力学。 近红外光谱。之前的结果已经确定了相对的 平衡溶液中的ATP、ADP和PO4的量可以是 用近红外光谱定量测定。的主要优势 使用近红外光谱是该方法的时间分辨能力， 允许光谱仪以非常快的速度监测反应 比例。考虑到这一点，我们观察到了ATP的水解 近红外光谱在十秒时间范围内。的变化 近红外光谱区反映了三磷酸腺苷被酶解成ADP的过程。 将这些光谱变化分离到它们各自的分量上并没有 已经完成；然而，光密度的总体变化 与无机磷的独立测定呈线性相关。使用 近红外技术的进步日新月异 时间分辨率的扩展以及时间分辨率的分离 组件是希望实现的。