Structure-Function Analysis of HィイD1+ィエD1-ATP Synthase by Solution and Solid State NMR

H-D1+H-D1-ATP 合成酶的溶液和固态 NMR 结构-功能分析

基本信息

  • 批准号:
    09480173
  • 负责人:
  • 金额:
    $ 8.06万
  • 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:
    1997
  • 资助国家:
    日本
  • 起止时间:
    1997 至 1999
  • 项目状态:
    已结题

项目摘要

The effect of nucleotide binding on the structure of the FィイD21ィエD2-ATPase βsubunit from thermophilic Bacillus PS-3 (TFィイD21ィエD2β) was investigated by monitoring the NMR signals of the twelve tyrosine residues. The 3,5-proton resonances of 12 tyrosine residues could be observed for the specifically deuterated β subunit. The assignment of 3,5-proton resonances of all the tyrosine residues was accomplished using fourteen mutant proteins, in each of which one or two tyrosine residues were substituted by phenylalanine. Binding of Mg・ATP induced an upfield shift of Tyr-341 resonance, suggesting that their aromatic rings are stacked to each other. Besides Tyr-341, the signal shift observed on Mg・ATP binding was restricted to the resonances of Tyr-148, Tyr-199, Tyr-238 and Tyr-307, suggesting that Mg・ATP induces a conformational change in the hinge region. This can be correlated to the change from the open to closed conformations as implicated in the crystal structure. Therefore, the intrinsic conformational change in the β subunit induced by the nucleotide binding is proposed to be one of the essential driving forces for the FィイD21ィエD2 rotation.A solid-state NMR spectrum for the correlation between anisotropic interactions of different spins depends on mutual orientations of anisotropic interactions, which specify dihedral angles. Thus, it can be used to determine the dihedral angle. This was applied to the ψand χ angles in alanine and valine, respectively. In the latter case, the bond lengths, bond angles also alter the spectrum. This enable us experimental determination of all structural parameters, which improves the accuracy of the dihedral angle determination.
通过监测12个酪氨酸残基的核磁共振信号,研究了核苷酸结合对嗜热芽孢杆菌PS-3 (TF γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ) atp酶β亚基结构的影响。12个酪氨酸残基的3,5-质子共振可以观察到特定氘化β亚基。使用14个突变蛋白完成了所有酪氨酸残基的3,5质子共振的分配,每个突变蛋白中有一个或两个酪氨酸残基被苯丙氨酸取代。Mg·ATP的结合诱导了tyr341共振的前场移位,表明它们的芳香环相互堆叠。除了tyr1 -341外,在Mg·ATP结合上观察到的信号移位仅限于tyr1 -148、tyr1 -199、tyr1 -238和tyr1 -307的共振,这表明Mg·ATP诱导了铰链区构象的变化。这可能与晶体结构中从开放构象到封闭构象的变化有关。因此,由核苷酸结合引起的β亚基的内在构象变化被认为是F _(_) _(_) _(_) _() _() _() _() _() _() _() _() _() _() _() _旋转的基本驱动力之一。不同自旋各向异性相互作用之间的相互关系取决于各向异性相互作用的相互取向,即指定的二面角。因此,它可以用来确定二面角。将此方法分别应用于丙氨酸和缬氨酸的ψ角和χ角。在后一种情况下,键长、键角也会改变光谱。这使得我们可以实验确定所有的结构参数,提高了二面角确定的精度。

项目成果

期刊论文数量(34)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
S.Kim: "The Interactions of ferric and ferrous cytochrome c with cardiolipin in phospholipid membranes studied by solid-state ^2H and ^<31>P NMR." J.Mol.Struct.441. 183-188 (1998)
S.Kim:“通过固态 2H 和 31 P NMR 研究了磷脂膜中铁和亚铁细胞色素 c 与心磷脂的相互作用。”
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    0
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T.Fujiwara: "Multidimensional solid-state nuclear magnetic resonance for correlating anisotropic interactions under magic-angle spinning conditions." J.Magn.Reson.124. 147-153 (1997)
T.Fujiwara:“多维固态核磁共振用于关联魔角旋转条件下的各向异性相互作用。”
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    0
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K.Noda: "Langmuir-Blodgett film of hydrogenase for electrochemical hydrogen production." Thin Solid Films. (印刷中).
K.Noda:“用于电化学制氢的朗缪尔-布洛杰特薄膜。”(正在出版)。
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    0
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S. Yoshioka, Y. Asao, S. Kojima, S. Sakurai, T. Fujiwara and H. Akutsu: "Molecular mobility of protein in lyophilized formulations linked to the molecular mobility of polymer excipients, as determined by high resolution solid-state ィイD113ィエD1C NMR"Pharmac
S. Yoshioka、Y. Asao、S. Kojima、S. Sakurai、T. Fujiwara 和 H. Akutsu:“通过高分辨率固态 D113D1C 测定,冻干制剂中蛋白质的分子迁移率与聚合物赋形剂的分子迁移率相关NMR"药学
  • DOI:
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  • 影响因子:
    0
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  • 通讯作者:
K. Ozawa, T. Meikari, K. Motohashi, M. Yoshida, and H. Akutsu: "Evidence for the presence of F-type ATPsynthase involved in sulfate-respiration in Desulfovibrio vulgaris"J. Bacteriol.. (in press).
K. Ozawa、T. Meikari、K. Motohashi、M. Yoshida 和 H. Akutsu:“存在参与脱硫弧菌硫酸盐呼吸的 F 型 ATP 合酶的证据”
  • DOI:
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  • 影响因子:
    0
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AKUTSU Hideo其他文献

AKUTSU Hideo的其他文献

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{{ truncateString('AKUTSU Hideo', 18)}}的其他基金

Molecular soft interactions regulating membrane-interface activities in living systems
调节生命系统膜界面活动的分子软相互作用
  • 批准号:
    15083101
  • 财政年份:
    2003
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Intermolecular soft interactions regulating H^+-ATPsynthase function
分子间软相互作用调节H^-ATP合酶功能
  • 批准号:
    15083203
  • 财政年份:
    2003
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Rotary catalysis mechanism of H^+-ATP synthase investigated by novel NMR methodology
通过新型 NMR 方法研究 H^-ATP 合酶的旋转催化机制
  • 批准号:
    14208082
  • 财政年份:
    2002
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Development of New Methodologies for Protein-Interaction Analysis
蛋白质相互作用分析新方法的开发
  • 批准号:
    10179101
  • 财政年份:
    1998
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas (A)
Structure and functions of a very small tetraheme protein -Molecular mechanism of bioremediation-
极小的四血红素蛋白的结构和功能-生物修复的分子机制-
  • 批准号:
    10044072
  • 财政年份:
    1998
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
DEVELOPMENT OF A BIOSUPRAMOLECULAR NMR SYSTEM WITH A SUPERCONDUCTING MAGNET
具有超导磁体的生物超分子核磁共振系统的开发
  • 批准号:
    03558029
  • 财政年份:
    1991
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research (B)
Interheme Electron Transfer in cytochrome C_3 and The Role of Aromatic Amino Acids.
细胞色素 C_3 中的组间电子转移和芳香族氨基酸的作用。
  • 批准号:
    01430024
  • 财政年份:
    1989
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (A)
Investigation on the dynamic structure of the lipid-containing virus PM2 and correlation with the infectivity.
含脂病毒PM2的动态结构及其与感染力的相关性研究
  • 批准号:
    60580221
  • 财政年份:
    1985
  • 资助金额:
    $ 8.06万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
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