Real-time observation of proton-pumping of membrane protein complexes using a novel high-sensitivity IR spectrometer

本项目旨在建立时间分辨红外光谱技术，实时观察生理条件下膜蛋白复合物的反应。采用飞秒红外激光器作为光源，构建了高灵敏度红外光谱仪。另一方面，开发了一种流动闪光测量系统来控制由笼状化合物的光解引起的反应。采用微通道流动电池，以最小的样品消耗获得光谱。将流动闪光系统安装在时间分辨紫外-可见吸收光谱仪上，对NO还原酶进行了测试。我们成功地以微秒级时间分辨率观察了该膜蛋白的酶促反应。流动闪光时间分辨红外光谱正在进行中。

This project aims to establish time-resolved IR spectroscopic techniques for observing reactions of membrane protein complexes under physiological conditions in real time. High-sensitivity IR spectrometer was constructed using a femtosecond IR laser as the light source. On the other hand, a flow-flash measurement system was developed to control reactions, which are triggered by the photolysis of caged compounds. Micro-channel flow cell was used to obtain a spectrum with minimal sample consumption. The flow-flash system was installed into time-resolved UV-visible absorption spectrometer and tested on NO reductase. We successfully observed the enzymatic reaction of this membrane protein with microsecond time resolution. Flow-flash time-resolved IR spectroscopy is ongoing.