Functional structures of nucleic acids and their recognition by proteins

核酸的功能结构及其蛋白质的识别

基本信息

  • 批准号:
    07458146
  • 负责人:
  • 金额:
    $ 3.84万
  • 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:
    1995
  • 资助国家:
    日本
  • 起止时间:
    1995 至 1996
  • 项目状态:
    已结题

项目摘要

A ribozyme in an RNA enzyme that can catalyze a biochemical reaction. Because a ribozyme requires a metal ions for a catalytic function, the ribozyme is considered as one of the metalloenzymes. Ribozyme reactions are very sensitive to species and concentration of metal ions, suggesting that divalent metal ions may play important roles for the chemical reaction and structural stabilization of the ribozyme.We have investigated the combined effects of metal ions on both the structural stabilization of the complex between a ribozyme (a leadzyme), CUGGGAGUCC,and a substrate, GGACCGAGCCAG,and the cleavage step in the active center of the complex has been also investigated kinetically.The results showed that the addition of Nd^<3+> in the presence of Pb^<2+> increased significantly the yield of the RNA cleavage reaction by the leadzyme, although other rare earth ions or divalent ions except Pb^<2+> did not promote the reaction. Further, kinetics for the leadzyme reaction was measured at various concentrations of Nd^<3+> and Pb^<2+>. At low concentration rations of Nd^<3+> under a constant total concentration of metal ions, Nd^<3+> increased the stablility of the complex between the leadzyme and the substrate. In contrast, at high concentration ratios of Nd^<3+>, the addition of Nd^<3+> decreased the stability of the complex. The rate constant of the cleavage step was maximized when the ratio of Nd^<3+> to Pb^<2+> was 1 : 1. These results suggest that the complex between the leadzyme and the substrate has binding sites for Nd^<3+> ion taht influence complex stability and catalyze directly the cleavage reaction. On the basis of the results, we propose a two-metal-ion mechanism in which Pb^<2+> and Nd^<3+> play the roles of base and acid catalyst, respectively.
RNA酶中的核酶,可以催化生化反应。由于核酶需要金属离子来发挥催化功能,因此核酶被认为是金属酶之一。核酶反应对金属离子的种类和浓度非常敏感,这表明二价金属离子可能对核酶的化学反应和结构稳定发挥重要作用。我们研究了金属离子对核酶(先导酶)CUGGGAGUCC和底物GGACCGAGCCAG之间复合物结构稳定性的综合影响,以及核酶中的裂解步骤 结果表明,在Pb^<2+>存在的情况下,添加Nd^<3+>显着提高了铅酶RNA裂解反应的产率,但除Pb^<2+>之外的其他稀土离子或二价离子并不促进该反应。此外,在不同浓度的Nd 3+ 和Pb 2+ 下测量了引导酶反应的动力学。在金属离子总浓度恒定的低Nd 3+ 浓度比下,Nd 3+ 增加了引导酶和底物之间的复合物的稳定性。相反,在高浓度比的Nd ^ 3+ 下,Nd ^ 3+ 的添加降低了络合物的稳定性。当 Nd^<3+> 与 Pb^<2+> 的比率为 1:1 时,裂解步骤的速率常数最大化。这些结果表明,引导酶和底物之间的复合物具有 Nd^<3+> 离子的结合位点,这会影响复合物的稳定性并直接催化裂解反应。基于这些结果,我们提出了一种双金属离子机理,其中Pb^<2+>和Nd^<3+>分别起碱催化剂和酸催化剂的作用。

项目成果

期刊论文数量(13)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Naoki Sugimoto: "Improved Thermodynamic Parameters and Helix Initiation Factor to Predict Stability of DNA Duplexes." Nucleic Acids Res.24. 4501-4505 (1996)
Naoki Sugimoto:“改进的热力学参数和螺旋起始因子可预测 DNA 双链体的稳定性。”
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    0
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P.C.Bevilacqua, N.Sugimoto, and D.H.Turner: "A Mechanistic Framework for the Second Step of Splicing Catalyzed by the Tetrahymena Ribozyme." Biochemistry. 35. 648-658 (1996)
P.C.Bevilacqua、N.Sugimoto 和 D.H.Turner:“四膜虫核酶催化剪接第二步的机制框架”。
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    0
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Naoki Sugimoto: "Effect of Metal Ions on DNA Ligation Catalyzed by a Deoxyribozyme." Nucleic Acids Res.,Symp.Series. 35. 191-192 (1996)
Naoki Sugimoto:“金属离子对脱氧核酶催化的 DNA 连接的影响。”
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  • 影响因子:
    0
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  • 通讯作者:
Naoki Sugimoto: "Energetics of Oligodeoxyribonucleotides with 5-methylcytosine" Rep. Prog. Poly. Phys. Jpn.38. 695-698 (1995)
Naoki Sugimoto:“寡脱氧核糖核苷酸与 5-甲基胞嘧啶的能量学”Rep. Prog。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Naoki Sugimoto: "The Stability of DNA and RNA G-Quartels." Nucleosides & Nucleotides. 15. 559-567 (1996)
Naoki Sugimoto:“DNA 和 RNA G-Quartels 的稳定性。”
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  • 影响因子:
    0
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SUGIMOTO Naoki其他文献

SUGIMOTO Naoki的其他文献

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{{ truncateString('SUGIMOTO Naoki', 18)}}的其他基金

Development of the system to change enzymatic functions without mutational approaches by using molecular crowding
开发利用分子拥挤无需突变方法即可改变酶功能的系统
  • 批准号:
    16K14041
  • 财政年份:
    2016
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Analyses of Protein Folding Codon affecting protein functions
蛋白质折叠密码子影响蛋白质功能的分析
  • 批准号:
    22655057
  • 财政年份:
    2010
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Development of monitoring technique using qNMR multivariate analysis for hazardous compounds in water
利用 qNMR 多变量分析监测水中有害化合物的技术的开发
  • 批准号:
    22590127
  • 财政年份:
    2010
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Development of novel nucleic acid materials that efficiently work in the intracellular molecular environment
开发在细胞内分子环境中有效工作的新型核酸材料
  • 批准号:
    21245040
  • 财政年份:
    2009
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Investigations of the molecular crowding effect on nucleic acid structures and development of nucleotide switches
分子拥挤效应对核酸结构和核苷酸开关开发的研究
  • 批准号:
    17350087
  • 财政年份:
    2005
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Evaluation of RNA cleavage reaction based on the prediction parameters for nucleic acid structures
基于核酸结构预测参数的RNA切割反应评估
  • 批准号:
    13132208
  • 财政年份:
    2001
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Database construction for the effect of metal ions on nucleic acid folding
金属离子对核酸折叠影响的数据库构建
  • 批准号:
    13480190
  • 财政年份:
    2001
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Development of novel gene-chips with unnatural nucleic acids
使用非天然核酸开发新型基因芯片
  • 批准号:
    12555231
  • 财政年份:
    2000
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Development of separation and detection of functional life molecules using combinatorial chemistry and ribozyme system
利用组合化学和核酶系统开发功能生命分子的分离和检测
  • 批准号:
    09440253
  • 财政年份:
    1997
  • 资助金额:
    $ 3.84万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
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