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Chemical Evolution of De novo Designed Artificial Proteins Incorporating Biological Functions

Research Project

Project/Area Number 10480153
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bioorganic chemistry
Research InstitutionKyushu Institute of Technology (1999)
Kyushu University (1998)

Principal Investigator

NISHINO Norikazu  Kyushu Institute of Technology, Faculty of Engineering, Professor, 工学部, 教授 (40145165)

Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1998: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsArtificial Protein / 4α-helix Bundle Structure / βαβαStructure / Flavin / Porphyrin-Fe(III) Complex / Peroxidase / βαβα錯体 / アルギニン / コラーゲン / 触媒機能 / 酵素モデル
Research Abstract

The chemical evolution of four-α-helix bundle polypeptide to a flavoenzyme model was attempted by designing single-chain 53-peptides. The flavoenzyme models were examined for catalytic oxidation reaction in aqueous solution. Since the hydrophobic core seemed to be too tightly aggregated for atalytic group, we tested a series of alkane-sulfonates to enlarge it by forming mixed micelles. The expanded hydrophobic core may more easily accommodate the hydrophobic substrate. The alkyl chain length longer than dodecyl enhanced the oxidation of benzyl-NAH by a flavoenzyme model by about 6-fold. These results suggest that the hydrophobic core in the bundle structure is useful to place the catalytic groups and have the advantage of accommodating hydrophobic substrates. On the other hand, a porphyrin derivative was incorporated into a single-chained two-α-helix polypeptide containing 29 amino acid residues via the thiol side chain of Cys residue. The dimerized Fe(III)-porphyrin-linked two-α-helix … More polypeptide was examined for the biomimetic peroxidase-like activity with HィイD22ィエD2 OィイD22ィエD2 or 3-chloroperbenzoic acid (mCPBA) as the oxidant. The kィイD2catィエD2/KィイD2MィエD2 value for the oxidation by the polypeptide with mCPBA was increased by 5000 times than that with HィイD22ィエD2OィイD22ィエD2.
Thus, we demonstrated the possibility to utilize the hydrophobic core of the four-α-helix bundle structure to evolve the artificial proteins chemically by de novo design to the artificial enzymes. Then, we chose a βαβα-structure as a polypeptide scaffold, since the βα-unit is found in TIM barrel. In order to place a porphyrin ring in the hydrophobic space in the folded structure, we designed a single-chained 49-peptide applying the amphiphilic α-helix and β-strand motifs. Peroxidase-like oxidation activity of this polypeptide occurred specifically in the presence of LPO rather than HィイD22ィエD2 OィイD22ィエD2 at low methanol content, reflecting that the hydrophobic inside was used as a binding site of the oxidant. We concluded that the de novo design of artificial proteins should be further polished to enhance their catalytic activities. Less

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] 富崎 欣也: "De Novo Design of Hemoprotein Model with Peroxidase Activity toward Lipophilic Peroxide"Chem. Lett.. 2000 (in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 富崎 欣也: "Chiral Assembly of A Pair of Free Base-Porphyrins and Peroxidase-like Activity of Iron(III)-Porphyrins in Four-α-Helix Bundle Structures with Dimerized Two-α-Helix Polypeptides"J. Chem. Soc., Perkin Trans. 2. 2000(in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 富崎 欣也: "Design and Characterization of Flavoenzyme Models in a Course of Chemical Evolution of Four-α-Helix Bundle Polypeptide"J. Chem. Soc., Perkin Trans. 2. 2000. 813-822 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 富崎 欣也: "Substrate Specificities of Artificial Flavo-Enzymes"Peptide Science- Present and future. 1999. 76-77 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 富崎 欣也: "Catalytic Activities of An Artificial Protein with Designed Loops"Peptide Science- Present and future. 1999. 78-79 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 金子 和明: "Catalytic Activities of An Artificial Heme Enzyme"Peptide Science- Present and future. 1999. 80-81 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Tomizaki: "De Novo Design of Heoprotein Model with Peroxidase Activity toward Lipophilic Peroxide"Chem. Lett.. 2000(in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Tomizaki: "Chiral Assembly of A pair of Free Base-Porphyrins and Peroxidase-like Activity of Iron(III)-porphyrins in Four-α-Helix Bundle Structures with Dimerized Two-α-Helix Polypeptides"J. Chem. Soc., Perkin Trans.. 2,2000(in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Tomizaki: "Design and Characterization of Flavoenzyme Models in a Course of Chemical Evolution of Four-α-Helix Bundle Polypeptide"J. Chem. Soc., Perkin Trans.. 2, 2000. 813-822 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Tomizaki: "Substrate Specificities of Artificial Flavo-Enzymes"Peptide Science-Present and Future. 76-77 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Tomizaki: "Catalytic Activities of An Artificial Protein with Designed Loops"Peptide Science-Present and Future. 78-79 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Kaneko: "Catalytic Activities of An Artificial Heme Enzyme"Peptide Science-Present and Future. 80-81 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 富崎欣也: "Design and characterization of flavoenzyme models in the course of chemical evolution of four-α-helix bundle polypeptides"J.Chem.Soc.,Perkin Trans.2.. 813-822 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 富崎欣也: "Chiral assembly of a pair of free base porphyrins and peroxidase-like activity of iron (III) porphyrins in four-α-helix bundle structures with dimerized two-α-helix polypeptides"J.Chem.Soc.,Perkin Trans.2.. in press. (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kazuaki Kaneko, Kin-ya Tomizaki, and Norikazu Nishino: "Catalytic activities of an artificial heme enzyme" Peptides Proceedings of the 1st International Peptide Symposium. in press. (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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