THERMODYNAMIC AND KINETIC STUDIES OF CYTOCHROME C OXIDASE

细胞色素C氧化酶的热力学和动力学研究

• 批准号: 3857972
• 负责人: R W HENDLER
• 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/3857972
• 关键词: Raman spectrometry; atomic absorption spectrometry; bacteriorhodopsins; bioenergetics; cell biology; cellular respiration; cytochrome b; cytochrome oxidase; oxidation reduction reaction; potentiometry; thermodynamics

A new continuous flow cell has been designed and built so that combined optical and Resonance raman spectroscopy can be used to monitor potentiometric titrations of cytochrome oxidase. In connection with these experiments, a new technique for controlling electrode voltages and/or currents has been developed. Use is made of computer I/O cards rather than analog circuit devices as used in the past. Software to accomplish this purpose, as well as to perform and monitor the titrations has been developed. In order to use the technique of singular value decomposition (SVD) to deconvolute kinetic spectra during the rapid turnover of cytochrome oxidase, a new ultra rapid spectrometer has been designed and built. The necessary software to control the instrument has been developed. This spectrometer will be capable of accumulating and storing sequential optical spectra with a time resolution of 10 (mu)sec. SVD has been used to analyze kinetic spectra obtained with bacteriorhodopsin, which was activated with different light intensities. We find that the light appears to "titrate" two centers which affect the kinetic properties of intermediates in the cycle.

设计和建造了一种新的连续流动池，以便结合 光学和共振拉曼光谱可用于监测 细胞色素氧化酶的电位滴定。 与这些相关 实验，一种控制电极电压和/或的新技术 电流已被开发。 使用计算机 I/O 卡而不是 过去使用的模拟电路设备。 实现此目的的软件 目的以及执行和监测滴定已 发达。 为了使用奇异值分解技术 （SVD）在快速周转期间对动力学谱进行解卷积 细胞色素氧化酶，一种新型超快速光谱仪已被设计和 建造的。 控制仪器所需的软件已 发达。 该光谱仪将能够积累和存储 时间分辨率为 10（μ）秒的连续光谱。 SVD 有 用于分析细菌视紫红质获得的动力学光谱， 被不同的光强度激活。 我们发现光 似乎“滴定”了两个影响动力学特性的中心 循环中的中间体。