1998 Fiscal Year Final Research Report Summary
Development of Novel Methods for Structural Analyses of Proteins
| Project/Area Number |
06276102
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Research Institution | Osaka University |
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Principal Investigator |
AIMOTO Saburo Inst. for Protein Research, Osaka Univ., Professor, 蛋白質研究所, 教授 (80029967)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Akira Tokyo Metropolitan Univ., Fac. of Sci., Associate Professor, 大学院・理学研究科, 助教授 (10183253)
SHIRAKAWA Masahiro Nara Inst. of Sci. and Tech., Dept. of Biosci., Associate Professor, バイオサイエンス研究科, 助教授 (00202119)
FUJIWARA Toshimichi Yokohama National Univ., Dept. of Engineering, Associate Professor, 工学部, 助教授 (20242381)
KATO Koichi Univ. of Tokyo School of Pharmaceutical Sci., Lecturer, 大学院・薬学研究科, 講師 (20211849)
TSUKIHARA Tomitake Inst. for Protein Research, Osaka Univ., Professor, 蛋白質研究所, 教授 (00032277)
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| Project Period (FY) |
1994 – 1997
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| Keywords | Protein / 3-dimensional Structure / Analytical Method / Protein Nucleic Acid Interaction / NMR / Solid-state NMR / Chemical Synthesis / X-ray Crystallography |
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| Research Abstract |
The members of this group focused their efforts on the development of novel methods for the analyses of protein structures and of their interactions. A new method was developed to elucidate the three-dimensional structure of unordered solid-state proteins such as proteins in a membrane by employing the solid-state NMR. A series of protein structures such as transcription factors in solution were determined by NMR through the basic methodological studies of solution NMR. A new method was developed based on a novel labeling method of a high molecular weight glycoprotein. Useful structural data of immunoglobulin G complex could be obtained by using the combination of the labeling method and NMR. An efficient method for the chemical preparation of proteins, including conjugated proteins such as phosphoproteins and glycoproteins, for structural studies was developed based on a thioester method. Synthetic method of stable-isotope labeled amino acids and bases in nucleic acids were established. Using a chemically synthesized labeled DNA, the structure of protein-DNA complex was determined by NMR. A novel technique to solve the crystal structures of super-macromolecules was developed, enabling us to increase the upper resolution limit of x-ray diffraction of Rice Dwarf virus crystals from 20A up to atomic resolution. In order to analyze the interaction between bio-macromolecules and aggregated structure of biomacromolecules, a system for x-ray small angle scattering, and a data collection method and a detector for neutron scattering for crystal structure analysis were developed. A new efficient algorithm to generate the structures of proteins and protein-DNA complexes was developed based on the NMR data.
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