DEVELOPMENT OF BIOPHYSICAL METHODS FOR STUDY OF BIO-MOLECULAR REACTIONS

研究生物分子反应的生物物理方法的发展

• 批准号: 3878933
• 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/3878933
• 关键词: adenosine diphosphate; adenosine triphosphate; antiseptic sterilization; blood /plasma substitute; calorimetry; chemical binding; chemical reaction; chlorine; hemoglobin; infrared spectrometry; macromolecule; spectrometry; temperature

The utilization of adenosine triphosphate as the prime energy source of many muscle and ion transport reactions is of considerable interest to the understanding of cellular processes. A major problem in studying the pre- steady state kinetics of these reactions has been the lack of ability to follow the disappearance of ATP and the appearance of the reaction products ADP and P04. We have recently utilized near infra-red spectroscopy to detect each of these compounds and have been able to make calibration curves for each in the 1 to 25 micromolar region. Many attributes of this system must be carefully controlled to allow us to have really good reproducibility and sensitivity i.e., approaching that of radiolabeled compounds which utilize P32 labelled ATP. Considerable progress has been made on the problem this year and we now know the measurements can be made. Laser Raman spectroscopy has been carried out on the P04 mixture at pH 11, 9, 6, and 1 as well as pH 7.4 where a mixture of the H2PO4, HP04 occurs but where we get the best spectral signature of ATP, ADP and P04. Much further work will be needed to elucidate the source of the NIR spectra and determine the conditions that affect the spectra. Work on the development of instrumentation for blood substitute continues but a number of technical problems has slowed down the completion of these instruments.

三磷酸腺苷作为主要能源的利用 许多肌肉和离子转运反应引起了人们的极大兴趣 了解细胞过程。前期研究中的一个主要问题 这些反应的稳态动力学一直缺乏能力 跟踪 ATP 的消失和反应产物的出现 ADP 和 P04。我们最近利用近红外光谱来 检测这些化合物中的每一种并能够进行校准 每个曲线在 1 至 25 微摩尔范围内。这个的很多属性 系统必须仔细控制才能让我们拥有真正好的 再现性和灵敏度，即接近放射性标记的再现性和灵敏度 利用 P32 标记的 ATP 的化合物。已经取得了长足的进步 今年针对这个问题进行了研究，现在我们知道可以进行测量了。 激光拉曼光谱已在 pH 11 下对 PO4 混合物进行， 9、6 和 1 以及 pH 7.4，其中出现 H2PO4、HP04 的混合物，但 我们获得了 ATP、ADP 和 P04 的最佳光谱特征。更进一步 需要开展工作来阐明近红外光谱的来源 确定影响光谱的条件。致力于开发工作 血液替代品仪器的使用仍在继续，但一些技术 问题减缓了这些文书的完成速度。