Time-Resolved Vibrational Spectroscopic Study of Higher Order Structural Changes of Protein Induced by Nanosecond Temperature Jump.
纳秒温度跳跃引起的蛋白质高阶结构变化的时间分辨振动光谱研究。
基本信息
- 批准号:10480187
- 负责人:
- 金额:$ 6.98万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B).
- 财政年份:1998
- 资助国家:日本
- 起止时间:1998 至 2000
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
A nanosecond temperature jump (T-jump) apparatus was constructed and combined with time-resolved Raman measurements to investigate thermal unfolding of a protein for the first time. The 1.56 μm-heat pulse with 9 ns width, 10 Hz was obtained through the two-step stimulated Raman scattering in D_2 gas involving seeding and amplification. To achieve uniform temperature rise, the counter-propagation geometry was adopted. The temperature rise was determined by anti-Stokes to Stokes intensity ratios of the 317 and 897 cm^<-1> bands of MoO^<2->_4 aqueous solution. The T-jump as large as 9℃ in 10 ns was attained. The unfolding of bovine pancreatic ribonuclease A was monitored with time-resolved Raman spectra excited at 532 nm. In the initial 200 ns after T-jump, the C-S stretch of Met residues exhibited 10% change of that expected from the steady state spectra and another 10% in 5 ms. The Raman intensity of S0^<2->_4 ions around 980 cm^<-1> increased at 100 μs delay and was shifted to a lower frequency than the steady state frequency of the elevated temperature, presumably due to partial release from the active site. The S-S stretches and tyrosine doublets displayed little change within 5 ms. Thus, the conformation change in the initial step of unfolding is not always concerted.
构建了纳秒温度跳变(T-jump)装置,并结合时间分辨拉曼测量首次研究了蛋白质的热展开。通过在D_2气体中进行两步受激拉曼散射,得到了宽度为9 ns、频率为10 Hz的1.56 μm热脉冲。为了实现均匀的温升,采用了反传播几何形状。温升由MoO^<2->_4水溶液的317和897 cm^<-1>波段的反Stokes与Stokes强度比决定。在10 ns内达到9℃的t跃。用532 nm激发的时间分辨拉曼光谱监测牛胰腺核糖核酸酶A的展开。在t跃迁后的最初200ns内,Met残基的C-S长度表现出稳态光谱预期值的10%的变化,在5ms内又表现出10%的变化。在980 cm^<-1>附近,S0^<2- bbb_4离子的拉曼强度在100 μs的延迟下增加,并转移到比高温稳态频率更低的频率,这可能是由于活性部位的部分释放所致。S-S链和酪氨酸双链在5 ms内变化不大。因此,展开初始阶段的构象变化并不总是一致的。
项目成果
期刊论文数量(15)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
K.Yamamoto et al.: "Construction of Novel Nanosecond Temperature Jump Apparatuses Applicable to Raman Mesurements and Direct Observation of Transient Temperature"Applied Spectroscopy. 54. 1591-1604 (2000)
K.Yamamoto 等人:“适用于拉曼测量和瞬态温度直接观察的新型纳秒温跃装置的构造”应用光谱学。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
K.Yamamoto, Y.Mizutani, and T.Kitagawa: "Constrctuion of novel nanosecond temperature jump apparatuses applicable to Raman measurements and direct observation of transient temperature."Applied Spectroscopy. 54. 1591-1604 (2000)
K.Yamamoto、Y.Mizutani 和 T.Kitakawa:“构建适用于拉曼测量和瞬态温度直接观察的新型纳秒温度跳跃装置。”应用光谱学。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
K.Yamamoto et al.: "Nanosecond Temperature Jump and Time-resolved Raman Study of Thermal Unfolding of Ribonuclease A"Biophysical Journal. 79. 485-495 (2000)
K.Yamamoto 等人:“核糖核酸酶 A 热展开的纳秒温度跳跃和时间分辨拉曼研究”生物物理学杂志。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
K.Yamamoto, Y.Mizutani, and T.Kitagawa: "Nanosecond temperature jump and time-resolved Raman study of thermal unfolding of ribonuclease A"Biophysical Journal. 79. 485-495 (2000)
K.Yamamoto、Y.Mizutani 和 T.Kitakawa:“核糖核酸酶 A 热展开的纳秒温度跳跃和时间分辨拉曼研究”生物物理学杂志。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
K. Yamamoto, Y. Mizutani and T. Kitagawa: "Nanosecond Temperature Jump and Time-resolved Raman Study of Thermal Unfolding of Ribonuclease A"Biophysical Journal. (in press). (2000)
K. Yamamoto、Y. Mizutani 和 T. Kitakawa:“核糖核酸酶 A 热展开的纳秒温度跳跃和时间分辨拉曼研究”生物物理学杂志。
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- 发表时间:
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- 影响因子:0
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KITAGAWA Teizo其他文献
KITAGAWA Teizo的其他文献
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{{ truncateString('KITAGAWA Teizo', 18)}}的其他基金
UV resonance Raman investigation on detection of higher order structural changes of heme proteins and elucidation of functional regulation mechanism
紫外共振拉曼研究检测血红素蛋白高阶结构变化并阐明功能调节机制
- 批准号:
24350086 - 财政年份:2012
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Structural Chemistry on Information Transduction through Allosteric Effects in Heme Proteins
通过血红素蛋白变构效应进行信息转导的结构化学
- 批准号:
21350098 - 财政年份:2009
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Structural Chemistry Involved in Discrimination of Diatomic Ligands and Transduction Mechanism of Sensed Information of Gas Sensing Heme Proteins
双原子配体识别的结构化学及气敏血红素蛋白传感信息的转导机制
- 批准号:
19350089 - 财政年份:2007
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
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 - 财政年份:2002
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Specially Promoted Research
Time-Resolved Vibrational Spectroscopic Investigation of Ultrafast Protein Dynamics Coupled with Photoreaction
超快蛋白质动力学与光反应耦合的时间分辨振动光谱研究
- 批准号:
12045264 - 财政年份:2000
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
Molecular Science on the Specific Roles of Metal Ions in Biological Functions.
金属离子在生物功能中特定作用的分子科学。
- 批准号:
08249105 - 财政年份:1996
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
Elucidation of Reaction Mechanisms of Biological Nitric Oxide by Vibrational Spectroscopy
振动光谱法阐明生物一氧化氮的反应机制
- 批准号:
07454157 - 财政年份:1995
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Detection of Fast Conformation Changes of Proteins by Time-Resolved Resonance Raman Spectroscopy
通过时间分辨共振拉曼光谱检测蛋白质的快速构象变化
- 批准号:
05453212 - 财政年份:1993
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Molecular Science for Elucidation of Proton Active Transport and Electron Transfers through Proteins.
通过蛋白质阐明质子主动传输和电子转移的分子科学。
- 批准号:
02453157 - 财政年份:1990
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Quantum Theory and Spectroscopy of Proteins as Biomachines
作为生物机器的蛋白质的量子理论和光谱学
- 批准号:
02305013 - 财政年份:1990
- 资助金额:
$ 6.98万 - 项目类别:
Grant-in-Aid for Co-operative Research (A)
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