2001 Fiscal Year Final Research Report Summary
Studies on molecular recognition mechanisms of biomolecules and their application
| Project/Area Number |
10145106
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (A)
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| Allocation Type | Single-year Grants |
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| Research Institution | Kyoto University |
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Principal Investigator |
IMANAKA Tadayuki Graduate School of Engineering, Kyoto University, Professor, 大学院・工学研究科, 教授 (30029219)
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| 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)
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| Project Period (FY) |
1998 – 2001
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| Keywords | Biomolecule / Recognition / Enzyme / DNA polymerase / Hvperthermophile / Chemotaxis / β-Propeller protein / Phytochelatin |
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| 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
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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
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