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: 3779525
• 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/3779525
• 关键词: adenosine diphosphate; adenosine triphosphate; bioengineering /biomedical engineering; biomedical equipment development; enzyme activity; enzyme substrate complex; infrared spectrometry; phosphorus

The study of ATP utilization by metabolic processes has been of considerable interest to the overall understanding of cellular activities. The most intriguing aspect of near infrared spectroscopy is the capability of monitoring a reaction on a very fast (millisecond) timescale. First of all, we must be able to accurately separate the reactants and products of ATP hydrolysis. We have employed near infrared spectroscopy to quantify ATP, ADP, and Pi and have successfully made calibration curves for each in the 1 to 25 micromolar regime. Recently, mixtures of the three phosphate compounds were also quantified in the same concentration region. Hence, the distinction of ATP, ADP, and Pi can be achieved with near infrared spectroscopy. Our next step has been to follow the hydrolysis of ATP both in the near infrared and with 31P coincidently. A necessary requirement for the success of near infrared spectroscopy is an accurate calibration. Once this calibration is attained, subsequent near infrared spectra can be compared and quantified with the calibrated model. On preliminary studies, percent errors of < 5% have been accomplished. With multiple linear regression techniques, a setting of six variables was found to yield the best fit to the data. This indicates that several factors may be involved in the hydrolysis of ATP, namely ATP-enzyme complexes and various other intermediates.

代谢过程中ATP利用的研究已经有很多