Cytochrome oxidase and proton translocation

1.为建立激光闪光光解技术测量组织氧浓度的系统提供基础数据，构建了快速扫描分光光度计、双闪光分光光度计和时间分辨吸收分光光度计。快速扫描和双闪光装置都是世界上性能最高的装置;前者允许以1 ms的最大时间分辨率记录200 nm跨度的512个连续吸收和反射光谱，后者的时间分辨率为1 μ s。这些设备已经广泛地用于相关领域的广泛领域中。其中一个显著的成就是发现了酵母黄素血红蛋白作为氧传感器的功能作用.利用该装置和光纤探头对肉中肌红蛋白的氧化过程进行了光谱跟踪。在通过解剖肉块而将内部暴露于空气氧时，切割表面的颜色从深棕色变为亮红色。光谱变化清楚地表明氧合肌红蛋白的形成，其特征在于在582和540 nm处的峰的增长，在591 nm处具有等吸光点。以591 nm为参比，在582 nm处的吸光度变化总是双相的.利用所建的装置研究了氧合血红蛋白的光解及其复合过程，并在不同氧浓度下收集了相关动力学数据。这些动力学参数与氧浓度相关，并建立了测量溶液中氧浓度的方法。基于这些数据，揭示了组织氧浓度测定中涉及的各种问题。

1. In order to collect fundamental data for establishing the system to measure the concentration of tissue oxygen by utilizing a laser flash photolysis technique, a rapid-scan spectrophotometer, double-flash spectrophotometer and a time-resolved absorption spectrophotometer were constructed. Both rapid-scan and double-flash apparatuses are of the highest performance ever built in the world; the former allows to record 512 consecutive absorption and reflectance spectra of a 200-nm span at the maximum time resolution of 1 ms, and the latter has a time resolution of 1mus. These apparatuses have been used extensively in wide areas of related fields. One of remarkable achievements is the finding of the functional role of yeast flavohemoglobin as an oxygen sensor.2. Oxygenation process of myoglobin contained in meat was followed spectrally by using the rapid-scan apparatus in combination with an optical probe made of optical fibers. Upon exposure of the interior to air oxygen by dissecting a meat lump, a color of the cut surface changes from dark brown to bright red. The spectral changes clearly indicate the formation of oxymyoglobin which is characterized by a growth of peaks at 582 and 540 nm with an isosbestic point at 591 nm. The absorbance changes followed at 582 nm against 591 nm as reference are invariably biphasic.3. The photodissociation of oxyhemoglobin and its recombination process have been studied by utilizing all of the apparatuses constructed and relevant kinetic data were collected at varying oxygen concentrations. These kinetic parameters are correlated with the oxygen concentration and the procedure to measure the oxygen concentration in solution has been established. Based on these data a variety of problems involved in the determination of tissue oxygen concentrations are revealed.