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Studies on molecular recognition mechanisms of biomolecules and their application

Research Project

Project/Area Number 10145106
Research Category

Grant-in-Aid for Scientific Research on Priority Areas (A)

Allocation TypeSingle-year Grants
Research InstitutionKyoto University

Principal Investigator

IMANAKA Tadayuki  Graduate School of Engineering, Kyoto University, Professor, 大学院・工学研究科, 教授 (30029219)

Co-Investigator(Kenkyū-buntansha) TAKAGI Masahiro  School of Materials Science, Japan Advanced Institute of Science and Technology, Professor, 材料科学研究科, 教授 (00183434)
SODE Koji  Faculty of Technology, Tokyo University of Agriculture & Technology, Associate Professor, 工学部, 助教授 (10187883)
OHTAKE Hisao  Department of Molecular Biotechnology, Hiroshima University, Professor, 工学部, 教授 (10127483)
Project Period (FY) 1998 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥112,600,000 (Direct Cost: ¥112,600,000)
Fiscal Year 2000: ¥36,500,000 (Direct Cost: ¥36,500,000)
Fiscal Year 1999: ¥39,000,000 (Direct Cost: ¥39,000,000)
Fiscal Year 1998: ¥37,100,000 (Direct Cost: ¥37,100,000)
KeywordsBiomolecule / Recognition / Enzyme / DNA polymerase / Hvperthermophile / Chemotaxis / β-Propeller protein / Phytochelatin / 分子認識 / Rubisco / DNA ligase / β-glycosidase / 基質特異性 / 始原菌 / バイオターゲティング / 環境応答分子 / 重金属捕捉ペプチド / phytochelatin / 抗体工学
Research Abstract

In cell surface, many recepter proteins are presented and the specific biomolecules are recognized by these recepter proteins. These mechanisms can be analyzed from chemical view points. To systemize the biotergeting, the recognition mechanism of these biomolecules were investigated. Dr. Imanaka studied mechanisms of molecular recognition for enzymes, and also discovered many enzymes with unusual molecular recognition (substrate and cofactor specificity). A hyperthermophilic archaeon, Thermococcus kodakaraensis KOD 1 was focused on. Detailed enzymatic analysis, along with structural analysis has been performed for various enzymes. Dr. Ohtake studied molecular mechanisms of chemotaxis in Pseudomonas aeruginosa. P. aeruginosa was attracted by 20 commonly occurring L-amino acids, organic acids, aromatic compounds, O_2, and inorganic phosphate (P_I). P. aeruginosa showed negative chemotactic responses toward thiocyanate and isothiocyanate esters, and volatile chlorinated aliphatic compound … More s. The methyl-accepting chemotaxis protein (MCP)-dependent pathway was analyzed in detail. Dr. Sode studied a novel strategy for the construction of proteins based on the molecular architecture of β-propeller protein (BPP). By multichimeric enzyme construction, the cumulative effect of functional regions of PQQGDH introduced in BPP was demonstrated. It was also demonstrated the W-motif exchange between two different BPP with similar scaffold, showing the future development of novel protein constructs based on BPP scaffolds. Dr. Takagi studied apoptosis caused by Cd^<2+> detoxification by a plant specific peptide, phytochelatin (Phytochelatin (PC) ; (gamma-Glu-Cys)n-Gly) as well as antioxidants (glutathione (GSH) and N-acetyl cysteine (NAC)). Antioxidants could detoxify Cd^<2+> toxicity that Cd^<2+> causes oxidative stress. Detoxification by PC was stronger than that by antioxidants and the effect seems to be synergistic with Bcl-2. Production of the plant specific PC in mammalian cells was attempted by transferring a gene for PC synthase from Arabidopsis dialiana to jurkat cell and transformed Jurkat cells produced PC and exhibited resistance to Cd^<2+>. Less

Report

(4 results)
  • 2001 Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Y.Hashimoto 他: "Extracellular synthesis, specific recognition, and intracellular degradation of cyclomaltodextrins by the hyperthermophilic archaeon Thermococcus sp. strain B 1001"J. Bacteriol. 183・17. 5050-5057 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K.Shiraki 他: "Comparative analyses of the conformational stability between a hyperthermophilic protein and its mesophilic counterpart"Eur. J. Biochem. 268・15. 4144-4150 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y.Kannan 他: "Active subtilisin-like protease from form a hyperthermophilic archaeon in a from with a putative prosequence"Appl Environ. Micro biol. 67・6. 2445-2452 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tajima, T 他: "Chemotaxis of the nematode Caenorhaditis elegans toward cycloheximide and quinine hydrochloride"J. Biosci. Bioeng. 91. 322-324 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitsutani, A 他: "Analysis of algicidal proteins of a diatom-lytic marine bacterium Pseudoalteromonas sp. strain A25 by two-dimentional electrophoresis"Phycologia. 40. 286-291 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] J.Okuda 他: "The Role of Conserved His775/781 in Membrane-Binding PQQ Glucose Dehydrogenase of Escherichia coli and Acinetobacter calcoaceticus"J. Biochem. Mol. Biol. Biophys. 4. 415-422 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y. Hashimoto et.al: "Extracellular synthesis, specific recognition, and intracellular degradation of cyclomaltodextrins by the hyperthermophilic archaeon Thermococcus sp. strain B 1001"J. Bacteriol. 183 ・ 17. 5050-5057 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Shiraki et.al: "Comparative analyzes of the conformational stability between a hyperthermophilic protein and its mesophilic counterpart"Eur.J. Biochem. 268 ・ 15. 4144-4150 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Y. Kannan et.al: "Active subtilisin-like protease from a hyperthermophilic archaeon in a form with a putative prosequence"Appl Environ. Microbiol. 67 ・ 6. 2445-2452 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tajima, T et.al: "Chemotaxis of the nematode Caenorhaditis elegans toward cycloheximide and quinine hydrochloride"J. Biosci. Bioeng. 91. 322-324 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitsutani, A et.al: "Analysis of algicidal proteins of a diatom-lytic marine bacterium Pseudoalteromonas sp. strain A25 by two-dimentional electrophoresis"Phycologja. 40. 286-291 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] J. Okuda et.al.: "The Role of Conserved His775/781 in Membrane-Binding PQQ Glucose Dehydrogenase of Escherichia coli and Acinetobacter calcoaceticus"J. Biochem. Mol. Biol. Biophys. 4. 415-422 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tadayuki Imanaka: "Presence of a structurally novel type Rubisco in the hyperthermophilic archaeon, Pyrococcus kodakaraensis KOD1."Journal of Biological Chemistry. 274(8). 5078-5082 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tadayuki Imanaka: "Rubisco from the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 is composed solely of large subunits and forms a pentagonal structure."Journal of Molecular Biology. 293(1). 57-66 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tadayuki Imanaka: "Rubisco from the hyperthermophilic archaeon, Thermococcus kodakaraensis."Methods in Enzymol.. 331. 353-365 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tadayuki Imanaka: "A DNA ligase from a hyperthermophilic archaeon with unique cofactor specificity."Journal of Bacteriology. 182(22). 6424-6433 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tadayuki Imanaka: "Gene analysis and enzymatic properties of thermostable β-glycosidase from Pyrococcus kodakaraensis KOD1."J.Bioscience and Bioengineering. 88(2). 130-135 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] S. Ezaki et al.: "Presence of a structurally novel type ribulose-bisphosphate carboxylase/oxygenase in the hyperthermophilic archaeon, Pyrococcus kodakaraensis KOD1"J. Biol. Chem.. 274. 5078-5082 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N. Rashid et al.: "Isolation and characterization of phychrotrophs from subterranean environments"J. Biosci. Bioeng.. 87. 746-751 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S. Ezaki et al.: "Gene analysis and enzymatic properties of thermostable β-glycosidase from Pyrococcus kodakaraensis KOD1"J. Biosci. Bioeng.. 88. 130-135 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S. Satofuka et al.: "Rapid method for detection and detoxification of heavy metal ions in water environments using phytochelatin"J. Biosci. Bioeng.. 88. 287-291 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N. Maeda et al.: "Ribulose bisphosphate carboxylase/oxygenase from the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 is composed solely of large subunits and forms a pentagonal structure"J. Mol. Biol.. 293. 57-66 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T. Tanaka et al.: "A unique chitinase with dual active sites and triple substrate binding sites from hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1"Appl. Environ. Microbiol.. 65. 5338-5344 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] R.N.Rahman et al.: "Sequence analysis of glutamate dehydrogenase (GDH) from the hyperthermophilic archaeon Pyrococcus sp.KOD1 and comparison of the enzymatic characteristics of native and recombinant GDHs." Mol.Gen.Genet.257. 338-347 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Haruki et al.: "Gene cloning and characterization of recombinant RNase HII from a hyperthermophilic archaeon." J.Bacteriol.180. 6207-6214 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Koga et al.: "Thermostable glycerol kinase from a hyperthermophilic archaeon: gene cloning and characterization of the recombinant enzyme." Protein.Eng.11. 1219-1227 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Nishioka et al.: "Characterization of two intein homing endonucleases encoded in the DNA polymerase gene of Pyrococcus kodakaraensis strain KOD1." Nucleic.Acids.Res.26. 4409-4412 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.A.Siddiqui et al.: "In vitro heat effect on heterooligomeric subunit assembly of thermostable indolepyruvate ferredoxin oxidoreductase." FEBS.Lett.4. 372-376 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] R.N.Rahman et al.: "Ion pairs involved in maintaining a thermostable structure of glutamate dehydrogenase from a hyperthermophilic archaeon." Biochem.Biophys.Res.Commun.30. 920-926 (1998)

    • Related Report
      1998 Annual Research Report

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

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