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.
核酶,核酶核糖核酸酶中能催化生化反应的核酶由于核酶需要金属离子的催化作用,所以核酶被认为是金属酶的一种。核酶反应对金属离子的种类和浓度非常敏感,提示二价金属离子可能在核酶的化学反应和结构稳定中起重要作用。我们研究了金属离子对核酶(引导酶)CUGGGAGUCC和底物GGACCGAGCCAG之间的络合物结构稳定的联合作用,并对络合物活性中心的切割步骤进行了动力学研究。结果表明,在Pb^&lt;2+和Gt;除Pb2+外,其他稀土离子或二价离子对铅酶的RNA切割反应没有促进作用,但能显著提高铅酶对RNA切割反应的产率。此外,还测定了不同浓度的Nd3+和Pb2+的铅酶反应动力学。在金属离子总浓度不变的情况下,低浓度的Nd3+与Gt;在金属离子总浓度不变的情况下,Nd3+增加了酶与底物之间络合物的稳定性。相反,在较高的Nd3+浓度比下,Nd3+的加入降低了络合物的稳定性。当Nd3+与Pb2+的比例为1:1时,切割步骤的速率常数最大。这些结果表明,酶与底物之间的络合物具有与Nd^&lt;3+&gt;离子的结合部位,从而影响络合物的稳定性,直接催化切割反应。在此基础上,提出了Pb2+和Nd3+分别充当碱催化剂和酸催化剂的双金属离子机理。

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。
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  • 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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