PHOSPHORESCENT-PROBED HEME PROTEINS IN AQUEOUS SOLUTION

磷光探测水溶液中的血红素蛋白

• 批准号: 3438572
• 负责人: MERLYN D. SCHUH
• 金额: $ 5万
• 依托单位: DAVIDSON COLLEGE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3438572
• 关键词: cytochrome c; cytochrome oxidase; diffusion; fluorescence spectrometry; fluorescent dye /probe; hemoglobin; hemoprotein; intermolecular interaction; laser spectrometry; myoglobin; phosphorescence; water solution

Understanding the dynamic internal motion of globular proteins is a problem of fundamental importance to biochemistry. However, it is difficult to follow and gauge the extent of such internal motion of proteins in solution. 2-bromo-6-naphthyl sulfate (BNS) has been found to phosphoresce in degassed aqueous solutions, and the phosphorescence is quenched specifically by the heme rings of heme proteins. By measuring the lifetime of BNS phosphorescence as a function of heme concentration, the rate constant for quenching, k/g, can be determined. Since the quenching of phosphorescence by heme proteins is diffusion controlled, the value of k/q can be used to estimate the extent to which dynamic motion of heme proteins makes the prosthetic heme rings accessible during contact with other molecules. BNS will be used to probe the accessibility of heme rings in myoglobin, cytochrome c, hemoglobin, cytochrome c peroxidase and other heme proteins. In addition to helping to better understand protein dynamics, it is anticipated that the proposed research will also give valuable insights into the functional mechanisms of heme proteins. Some of the most important physiological functions of living systems are performed by heme proteins (e.g. transport of electrons in the production of metabolic energy and transport and storage of O/2 for oxidation of foods). A better understanding of the mechanisms of these proteins will eventually allow the development of ways to control for better health the physiological functions determined by heme proteins. An ultimate long-term objective will be to develop and use BNS as a phosphorescent probe of other specific biological polymers in other solvent environments, such as natural or artificial membranes.

了解球状蛋白质的动态内部运动是 一个对生物化学至关重要的问题。 然而，在这方面， 很难跟踪和衡量这种内部的程度， 蛋白质在溶液中的运动 2-溴-6-萘基硫酸盐（BNS） 已发现在脱气水溶液中发磷光， 磷光被血红素环特异性地猝灭， 血红素蛋白 通过测量BNS的寿命 磷光作为血红素浓度的函数， 可以确定淬火常数k/g。 以来 血红素蛋白对磷光的猝灭是扩散 控制，k/q的值可以用来估计的程度， 血红素蛋白质的动态运动使得假肢 血红素环在与其他分子接触时可接近。 BNS将用于探测血红素环的可及性， 肌红蛋白，细胞色素c，血红蛋白，细胞色素c过氧化物酶 和其他血红素蛋白。 除了帮助更好地 了解蛋白质动力学，预计提出的 研究还将提供有价值的见解， 血红素蛋白的机制。 一些最重要的 生命系统的生理功能是由血红素完成的 蛋白质（例如，在生产过程中的电子传输） 代谢能量和运输和储存用于氧化的O/2 食物）。 更好地了解这些机制 蛋白质最终将允许开发控制 为了更好的健康，由血红素决定的生理功能 proteins. 一个最终的长期目标将是开发和使用BNS， 其他特定生物聚合物的磷光探针， 其它溶剂环境，例如天然或人工的 膜。