Development of Detection Methods of Dynamical Higher Order Structures of Proteins and Its Application to Elucidation of Structure-Function Relations of Proteins.

继人类基因组问题之后，结构生物学成为生物物理化学的主干学科，“蛋白质3000”项目已成功开展。本项目旨在利用x射线晶体学和核磁共振光谱揭示蛋白质结构。因此，许多蛋白质的静态结构已经解决，但动态结构仍有待揭示，以了解蛋白质如何执行其功能。我们的研究旨在提供更详细的大型蛋白质的活性位点结构和其功能状态下的动力学结构。为此，采用了振动光谱学。开发了获得时间分辨可见光和紫外共振拉曼光谱和显微镜FTIR光谱的新系统，并应用于以下8个对象；1)利用皮秒时间分辨可见光共振拉曼光谱检测发色团的超快结构变化；2)利用亚纳秒时间分辨紫外共振拉曼光谱检测p…More蛋白质高阶结构的快速变化；3)利用激光温度跳变技术阐明蛋白质折叠/展开的主要过程；5)阐明DNA光解酶光诱导DNA修复的主要过程，6)淀粉样纤维结构的FTIR显微光谱研究及其形成的触发机制，7)气体传感血红素蛋白对环境的感知和信息转导到功能域的机制，8)血红素酶对氧激活和质子主动转运的机制。在实践中，我们已经构建了几个创造性的时间分辨共振拉曼系统，涵盖了广泛的激发波长（从紫外到近红外）和时间分辨率（从皮秒到秒）。我们成功地将这些系统应用于血红蛋白和肌红蛋白等基本蛋白质以及一些新的蛋白质，如气体传感器蛋白质。我们成功地阐明了气体传感器蛋白的靶标识别和信息传递机制。少

Structural biology became a main subject in biophysical chemistry after the human genome problem, and 'Protein 3000' project has been successfully carried out. This project intended to uncover the protein structures with x-ray crystallography and NMR spectroscopy. Thus, the static structures of many proteins have been solved, but dynamical structures remained to be revealed to understand how proteins perform their functions. Our research aimed to provide more detailed structures of active sites of large proteins and also dynamical structures in their functioning state. For this purpose vibrational spectroscopy was adopted. New systems for obtaining time-resolved visible and ultraviolet resonance Raman spectra and microscope FTIR spectra were developed and applied to the following eights subjects; 1) Detection of ultrafast structural change of chromophores by using picosecond time-resolved visible resonance Raman spectroscopy, 2) Detection of fast changes of higher order structures of p … More roteins by using subnanosecond time-resolved ultraviolet resonance Raman spectroscopy, 3) Elucidation of the primary process in protein folding/unfolding by using laser temperature jump technique, 4) Development of a novel spectroscopy for functionally-important protein fluctuation, 5) Elucidation of the primary process in the light-induced DNA repair by DNA photolyases, 6) FTIR microspectroscopic investigation of amyloid fibril structures and a trigger mechanism along its formation, 7) Mechanism of sensing of environment and information transduction to a functional domain by gas sensor heme proteins, 8) Elucidation of mechanism of oxygen activation and proton active transport by heme enzymes. In practice we have constructed several inventive time-resolved resonance Raman systems, which cover a wide ranges of excitation wavelengths (from ultraviolet to near infrared) and time-resolution (from picosecond to second). We applied these systems successfully to basic proteins like hemoglobin and myoglobin and some new proteins like gas sensor proteins. We succeeded in elucidating mechanisms of target discrimination and information transmission in the gas sensor proteins. Less