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Development of separation and detection of functional life molecules using combinatorial chemistry and ribozyme system

Research Project

Project/Area Number 09440253
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field 分離・精製・検出法
Research InstitutionKonan University

Principal Investigator

SUGIMOTO Naoki  Department of Chemistry, Konan University Professor, 理学部, 教授 (60206430)

Project Period (FY) 1997 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1998: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1997: ¥3,900,000 (Direct Cost: ¥3,900,000)
KeywordsCombinatorial chemistry / Ribozyme / Deoxyribozyme / RNA cleavage / Calcium ion / Metal ion / DNA chip / Avidin- biotin interaction / リポザイム / RNA切断活性 / DNA / RNA / 反応速度 / 分子認識 / ペプチド / コンビナトリアルケミストリー / インビトロセレクション / RecA / 機能性材料 / ファージディスプレイ / 遺伝子診断 / 核酸 / 希土類イオン / 反応メカニズム / 機能性核酸分子
Research Abstract

Catalytic DNA is a promising class of nucleic acid enzyme for possible use as a therapeutic agent and is also interesting in comparison with catalytic RNAs. In this study, we investigated the effect of metal ions and sequence on an original deoxyribozyme, d(GCCTGGCAG_1 G_2 C_3 T_4 A_5 G_6 C_7 T_8 A_9 C_<10> A_<11> A_<12> C_<13> G_<14> A_<15> GTCCCT), which binds to an RNA substrate, r(GAAGACA↓UGCCAGCG), cleaving the RNA substrate at one site indicated by the arrow. The results show that the ability of metal ions to promote the RNA cleavage reaction by the original deoxyribozyme is Mn^<2+> >Mg^<2+> >Ca^<2+> * Ba^<2+>. This result is very similar to the previous tendency observed in the case of a hammerhead ribozyme. Thus, these results suggest that RNA cleavage by the deoxyribozyme dependence on metal ions for catalysis is similar to that with ribozymes. On the other hand, a nucleotide deletion from the active domain of the original deoxyribozyme results in a novel deoxyribozyme with high Ca^<2+> -dependency, a situation which is not observed with the original deoxyribozyme or hammerhead ribozymes.

Report

(5 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (38 results)

All Other

All Publications (38 results)

  • [Publications] Naoki Sugimoto: "Thermodynamics-Structure Relationship of Single Mismatches in RNA/DNA Duplexes"Biochemistry. 39. 11270-11281 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tatsuo Ohmichi: "Kinetic Property of Bulged Helix Formation : Analysis of Kinetic Behavior Using Nearest-Neighbor Parameters"J.Am.Chem.Soc.. 122. 11286-11294 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Peng Wu: "Transition characteristics and thermodynamic analysis of DNA duplex formation : a quantitative consideration for the extent of duplex association"Nucleic Acids Res.. 28. 4762-4768 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yasuhide Okumoto: "Effects of metal ions and catalytic loop sequences on the complex formation of a deoxyribozyme and its RNA substrate"J.Inorg.Biochem.. 82. 189-195 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Junji Kawakami: "In vitro selection of aptamers that act with Zn^<2+>"J.Inorg.Biochem.. 82. 197-206 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Jin Zou: "A role of the Trp-His interaction in the conformational switch between α-helix and β-sheet in short alanine-based peptides"J.Chem.Soc.,Perkin Trans.2. 2135-2140 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 杉本直己: "ヒトゲノム最前線 21世紀は遺伝子の時代ゲノム化学とSNPs解析 化学の果たす役割。"化学同人. 27 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Sugimoto, M.Nakano, and S.Nakano: "Thermodynamics-Structure Relationship of Single Mismatches in RNA/DNA Duplexes"Biochemistry. 39. 11270-11281 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Ohmichi, H.Nakamuta, K.Yasuda, and N.Sugimoto: "Kinetic Property of Bulged Helix Formation : Analysis of Kinetic Behavior Using Nearest-Neighbor Parameters"J.Am.Chem.Soc.. 122. 11286-11294 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] P.Wu and N.Sugimoto: "Transition characteristics and thermodynamic analysis of DNA duplex formation : a quantitative consideration for the duplex association"Nucleic Acids Res. 28. 4762-4768 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Okumoto and N.Sugimoto: "Effect of metal ions and sequences in the catalytic loop on complex formation between a catalytic DNA with RNA cleavage activity and its RNA substrate"J.Inorg. Biochem. 82. 189-195 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Kawakami, H.Imanaka, Y.Yokota, and N.Sugimoto: "In Vitro Selection of Aptamers That Act with Zn2+"J.Inorg.Biochem. 82. 197-206 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] J.Zou and N.Sugimoto: "A role of Trp-His interaction in the conformational switch between a-helix and b-sheet in short alanine- based peptides"J.Chem.Soc., Perkin Trans.. 2. 2135-2140 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Naoki Sugimoto: "Thermodynamics-Structure Relationship of Single Mismatches in RNA/DNA Duplexes"Biochemistry. 39. 11270-11281 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tatsuo Ohmichi: "Kinetic Property of Bulged Helix Formation : Analysis of Kinetic Behavior Using Nearest-Neighbor Parameters"J.Am.Chem.Soc.. 122. 11286-11294 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Peng Wu: "Transition characteristics and thermodynamic analysis of DNA duplex formation : a quantitative consideration for the extent of duplex association"Nucleic Acids Res.. 28. 4762-4768 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yasuhide Okumoto: "Effects of metal ions and catalytic loop sequences on the complex formation of a deoxyribozyme and its RNA substrate"J.Inorg.Biochem.. 82. 189-195 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Junji Kawakami: "In vitro selection of aptamers that act with Zn^<2+>"J.Inorg.Biochem.. 82. 197-206 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Jin Zou: "A role of the Trp-His interaction in the conformational switch between α-helix and β-sheet in short alanine-based peptides"J.Chem.Soc.,Perkin Trans.2. 2135-2140 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 杉本直己: "ヒトゲノム最前線-21世紀は遺伝子の時代 ゲノム化学とSNPs解析 化学の果たす役割"化学同人. 27 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Sugimoto: "Effect of metal ions and sequence of deoxyribozymes on their RNA cleavage activity"J.Chem.Soc.,Perkin Trans.2. 1381-1386 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Sugimoto: "A novel screening method for combinatorial chemistry for low affinity interactions"Chem.Commun.. 677-678 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] J.Kawakami: "A selection of short peptides that interact with a porphyrin as a small target by immobilized phage display"Chem.Commun.. 1765-1766 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Sugimoto: "α-β structural Transition of Short Oligopeptides by Water/Organic Solvent Titration"Chem.Lett.. 637-638 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Nakano: "Nucleic Acid Duplex Stability : Influenece of Base Composition on Cation Effects"Nucleic Acids Res.. 27. 2957-2965 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] J.Zou: "Effect of copper ion on conformations of short alanine-based peptides with single Trp/His stacking interactions"Protein and Peptide Lett.. 6. 373-378 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Naoki Sugimoto: "Effect of Cu^<2+> on the Complex Formation between a Deoxyribozyme and its Substrates" Nucleosides & Nucleotides. 17. 565-574 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Junji Kawakami: "In Vitro Selection of Aptamers that Recognize a Monosaccharide" Analytica Chimica Acta. 365. 95-100 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Naoki Sugimoto: "Reaction Field for Efficient Porphyrin Metallation Catalysis Producedby Self-assembly of a Short DNA Oligonucleotide" Chemical Communications. 1533-1535 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shu-ichi Nakano: "Single-stranded DNA Recognition of a 24-mer Peptide Derived from RecA Protein" Bulletin of the Chemical Society of Japan. 71. 2205-2210 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Naoki Sugimoto: "Effect of Cation Concentration on Duplex Stability of Nucleic Acids with Unnatural Base Pairs" Nucleic Acids Res.,Symp.Series. 39. 121-122 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tatsuo Ohmichi: "Effect of Substrate RNA Sequence on the Cleavage Reaction by a Short Ribozyme" Nucleic Acids Res. 26. 5655-5661 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tatsuo Ohmichi: "Role of Nd^<3+> and Pb^<2+> on the RNA Cleavage Reaction by a Small Ribozyme" Biochemistry. 36. 3514-3521 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoki Sugimoto: "Experimental Quantitation of Cation Effect on Nucleic Acid Stability" Nucleic Acids Res., Symp. Series. 37. 147-148 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoki Sugimoto: "Stabilities of Internal rU-dG and rG-dT pairs in RNA/DNA Hybrids" Nucleic Acids Res., Symp. Series. 37. 199-200 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoki Sugimoto: "A 24mer Peptide Derived from RecA Protein Can Discriminate a Single-Stranded DNA from a Double-Stranded DNA" Chemical Communications. 2125-2126 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoki Sugimoto: "Sandwiching Interaction of Peptides with a Porphyrin" Chemistry Letters. 939-940 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Naoki Sugimoto: "Roles of 2'-Hydroxyls of Leadzyme on an RNA cleavage" Bulletin of the Chemical Society of Japan. 70. 2743-2747 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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