Functionally-important protein dynamics of photosensor proteins as revealed by time-resolved resonance Raman spectroscopy

时间分辨共振拉曼光谱揭示光传感器蛋白的功能重要的蛋白质动力学

• 批准号: 17350009
• 负责人: MIZUTANI Yasuhisa
• 金额: $ 10.14万
• 依托单位: Osaka University (2006-2007)Kobe University (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17350009/
• 关键词: resonance Raman spectroscopy; time-resolved spectroscopy; rotein dynamics; イエロープロテイン; センサータンパク質; ミオグロビン

Functionally-important protein dynamics of some sensor proteins have been studied by time-resolved resonance Raman spectroscopy. Chief results are as follows.(1) Construction of a time-resolved ultraviolet resonance Raman spectrometer(2) Picosecond Protein Response to the Chromophore Isomerization of Photoactive Yellow Protein (PIP)Picosecond time-resolved ultraviolet resonance Raman (UVRR) spectra of PYP were measured. UVRR bands attributed to the vibration of tyrosine and tryptophan residues showed a spectral change upon photoreaction. It was found that the hydrogen-bond strength between the chromophore and Y42 increases in the pG* state. The ultrafast change in the tryptophan band revealed that a photo-induced structural change of the chromophore had propagated to the W119 region, located 12 A from the chromophore, within picoseconds.(3) Structural Dynamics of FixLon Signal Transduction and Ligand Discrimination FixLFixL is a heme-based O2 sensor protein, which responds to low O2 co … More ncentration by activating the transcriptional activator FixJ. Signal transduction is initiated by the dissociation of O2 from the sensor domain of FixL, resulting in protein conformational changes that are transmitted to a histidine kinase domain. To gain insight into the FixL sensing mechanism, we monitored changes in the protein's structure in the picosecond to millisecond timeframe, following the dissociation of the ligand using time-resolved resonance Raman spectroscopy. The FixL spectra show that there are three steps in the dynamic structural changes. Ligand-dependent structural dynamics are observed in the earliest step. Based on comparisons of these structural changes, a scheme for the signal transduction of FixL is proposed. Similar spectral changes were observed both for the sensor domain and for the full length protein, although structural changes occurred faster with the former than with the latter. This difference in rate suggests that the structural changes occurring in the heme pocket are coupled to those of the kinase domain. Less

利用时间分辨共振拉曼光谱研究了一些传感蛋白质的功能动力学。主要结果如下。(1)时间分辨紫外拉曼共振光谱仪的研制（2）光敏黄蛋白（PIP）对生色团异构化的皮秒响应测量了PYP的皮秒时间分辨紫外拉曼共振（UVRR）光谱。UVRR带归因于酪氨酸和色氨酸残基的振动显示了光反应后的光谱变化。结果发现，生色团和Y42之间的氢键强度增加，在pG* 状态。超快变化的色氨酸带显示，光诱导的结构变化的发色团已经传播到W119区域，位于12 A的发色团，在皮秒。(3)FixLon信号转导和配体识别的结构动力学研究FixLFixL是一种基于血红素的氧传感器蛋白，它对低浓度的O2产生响应， ...更多信息 通过激活转录激活因子FixJ进行富集。信号转导是由O2从FixL的传感器结构域解离而引发的，导致蛋白质构象变化，并将其传递到组氨酸激酶结构域。为了深入了解FixL传感机制，我们使用时间分辨共振拉曼光谱法在配体解离后，在皮秒至毫秒的时间范围内监测蛋白质结构的变化。FixL光谱表明，动态结构变化有三个步骤。配体依赖的结构动力学观察在最早的步骤。基于这些结构变化的比较，FixL的信号转导的方案提出。类似的光谱变化，观察传感器域和全长蛋白质，虽然结构变化发生更快，前者比后者。这种速率的差异表明血红素口袋中发生的结构变化与激酶结构域的结构变化相耦合。少

项目成果

時間分解共鳴ラマン分光法によるタンパク質ダイナミクスの観測:構造変化と機能

通过时间分辨共振拉曼光谱观察蛋白质动力学：结构变化和功能

• 发表时间: 2007
• 作者: T. Odagiri;M. Murata;K. Funatsu;T. Tanabe;I. H. Suuki;M. Kitajima;and N. Kouchi;水谷泰久
• 通讯作者: 水谷泰久

The formation of hydrogen bond in the proximal heme pocket ofHemAT-Bs upon ligand binding

配体结合后 HemAT-B 近端血红素袋中氢键的形成

• 发表时间: 2007
• 期刊: Biochem.Biophys.Res.Commun 357
• 作者: Hirokazu IWAWAKI;Isao SINOHARA;Toshikazu KATO;Hideaki Yoshimura
• 通讯作者: Hideaki Yoshimura

Picosecond Protein Response to the Chromophore Isomerization of Photoactive Yellow Protein: Selective Observation of Tyrosine and Tryptophan Residues by Time-Resolved Ultraviolet Resonance Raman Spectroscopy

皮秒蛋白对光敏黄色蛋白发色团异构化的响应：通过时间分辨紫外共振拉曼光谱选择性观察酪氨酸和色氨酸残基

• 发表时间: 2007
• 期刊: J. Phys. Chem. B 111
• 作者: R.Katai;S.Morita;M.Goto;Misao Mizuno
• 通讯作者: Misao Mizuno

ピコ秒時間分解紫外共鳴ラマン分光によるタンパク質二次構造の高速変化の観測

利用皮秒时间分辨紫外共振拉曼光谱观察蛋白质二级结构的快速变化

• 发表时间: 2007
• 作者: M. Murata;T. Odagiri and N. Kouchi;董春鳳;水野操
• 通讯作者: 水野操

Primary protein response after ligand photodissociation in carbonmonoxy myoglobin

• DOI: 10.1073/pnas.0611560104
• 发表时间: 2007-06-05
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Sato, Akira;Gao, Ying;Mizutani, Yasuhisa
• 通讯作者: Mizutani, Yasuhisa