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

蛋白质动态高阶结构检测方法的发展及其在阐明蛋白质结构功能关系中的应用。

基本信息

批准号：
14001004
负责人：
KITAGAWA Teizo
金额：
$ 292.86万
依托单位：
Institute Molecular Science (2006)National Institutes of Natural Sciences Okazaki Research Facilities (2004-2005)Okazaki National Research Institutes (2002-2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Specially Promoted Research
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2006
项目状态：
已结题

项目摘要

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

人类基因组问题之后，结构生物学成为生物物理化学的一个主要学科，“蛋白质3000”计划已成功开展，该计划旨在通过X射线晶体学和核磁共振波谱揭示蛋白质的静态结构。许多蛋白质已经得到解决，但动态结构仍有待揭示，以了解蛋白质如何发挥其功能，我们的研究旨在提供大蛋白质活性位点的更详细结构以及其功能状态下的动态结构。采用了获得时间分辨可见光和紫外共振拉曼光谱以及显微镜FTIR光谱的新系统，并将其应用于以下八个课题：1）利用皮秒时间分辨可见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检测蛋白质高阶结构的快速变化光谱学，3）利用激光温度跳跃技术阐明蛋白质折叠/展开的主要过程，4）开发用于功能重要的蛋白质波动的新型光谱学，5）通过激光温度跳跃技术阐明光诱导DNA修复的主要过程DNA 光解酶，6) 淀粉样原纤维结构及其形成过程中的触发机制的 FTIR 显微光谱研究，7) 环境感知和信息转导到功能域的机制通过气体传感器血红素蛋白，8）阐明血红素酶的氧活化和质子主动运输机制在实践中，我们构建了几种创造性的时间分辨共振拉曼系统，其涵盖了广泛的激发波长（从紫外线到近红外线）。）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气体传感器蛋白质等一些新蛋白质。阐明气体传感器蛋白质中的目标辨别和信息传输机制。