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。

• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14001004/
• 关键词: Resonance Raman; Gas Sensor Heme Protein; Higher Order Structures; Ultraviolet Resonance Raman; FTIR Microscopy; Amvloid; Time-resolved Raman; Vibrational Spectroscopy; 時間今解共鳴ラマン; 蛋白質動的構造; 時間分解振動分光; センサー蛋白質; 温度ジャンプ; DNA結合蛋白; 赤外線分光; DNAフォトリア

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

人类基因组问题后，结构生物学成为生物物理化学的一个重要研究方向，“蛋白质3000”计划已经成功实施。该项目旨在用X射线晶体学和NMR光谱学揭示蛋白质结构。因此，许多蛋白质的静态结构已经得到解决，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蛋白质活性位点的结构，以及它们在功能状态下的动力学结构。为此，采用了振动光谱。本文介绍了一种新的时间分辨可见光和紫外共振拉曼光谱及显微傅里叶变换红外光谱系统，并将其应用于以下八个方面：1）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）皮秒时间分辨可见光共振拉曼光谱检测发色团的高阶结构的快速变化; ...更多信息 利用亚纳秒时间分辨紫外共振拉曼光谱技术研究鱼藤蛋白，3）利用激光温度跳跃技术阐明蛋白质折叠/去折叠的主要过程，4）开发一种新的功能重要的蛋白质波动光谱，5）阐明DNA光解酶光诱导DNA修复的主要过程，6）淀粉样蛋白原纤维结构的FTIR显微光谱研究和沿着其形成的触发机制，7）通过气体传感器血红素蛋白对环境的传感和信息转导到功能域的机制，8）阐明血红素酶的氧活化和质子主动转运机制。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，其覆盖了宽范围的激发波长（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碱性蛋白质如血红蛋白和肌红蛋白以及一些新的蛋白质如气体传感器蛋白。我们成功地阐明了气体传感器蛋白的目标识别和信息传递机制。少

项目成果

Stopped‐flow spectrophotometric and resonance Raman analyses of aldoxime dehydratase involved in carbon–nitrogen triple bond synthesis

碳氮三键合成中醛肟脱水酶的停流分光光度法和共振拉曼分析

• DOI: 10.1016/j.febslet.2005.01.037
• 发表时间: 2005
• 期刊: FEBS Letters
• 影响因子: 3.5
• 作者: Ken;H. Kumita;T. Ohta;K. Konishi;Y. Hashimoto;H. Higashibata;T. Kitagawa;Y. Shiro;Michihiko Kobayashi
• 通讯作者: Michihiko Kobayashi

"Resonance Raman Spectroscopy" J. Porphyrins Phthalocyanines

《共振拉曼光谱》J. 卟啉 酞菁

• 发表时间: 2002
• 期刊: Society of Porphyrine and Phtalocyanines
• 作者: Kitagawa;T.
• 通讯作者: T.

化学と工業「気体センサー蛋白質が特異的に気体分子を認識し、応答するメカニズム」

化学与工业“气体传感器蛋白特异性识别和响应气体分子的机制”

• 发表时间: 2007
• 作者: Nakamura;Itaru;内田 毅
• 通讯作者: 内田 毅

ラマン分光(光科学研究の最前線)

拉曼光谱（光学科学研究的前沿）

• 发表时间: 2005
• 作者: Kamijo;Shin;内田 毅
• 通讯作者: 内田 毅

「時間分解振動分光法で観たタンパク質の動き」ナノバイオテクノロジーの最前線

“通过时间分辨振动光谱观察蛋白质的运动”纳米生物技术的最前沿

• 发表时间: 2003
• 作者: Kamijo;Shin;J.M.STEVENS;Asano Toru;Asano Toru;T.OHTA;H.HIRAMATSU;R. Hamasaki;内田 毅;A. I. Siriwardana;N. Tsuboya;内田 毅;I. Kadota;N. Asao;水谷 泰久
• 通讯作者: 水谷 泰久