| Co-Investigator(Kenkyū-buntansha) |
FUJITO Teruaki JEOL,Service head office, section chief, 分析機器サービス本部, 課長(研究職)
KUBO Atsushi Kyoto Univ., Graduate School of Sci., Instructor, 大学院・理学研究科, 助手 (20205115)
TAKEGOSHI Kiyonori Kyoto Univ., Graduate School of Sci., Asso.Prof., 大学院・理学研究科, 助教授 (10206964)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 1997: ¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1996: ¥8,400,000 (Direct Cost: ¥8,400,000)
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| Research Abstract |
This study attempted to develop methods for precise and accurate determination of carbon-carbon and carbon-nitrogen distances as well as dihedral angles phi, psi in a particular local region of a biomolecule. It was aimed at applying to structural problems in a biology which have yet not been solved by either X-ray crystallography or solution-state NMR.
In this year, we elaborated several methods which we had developed, and applied them to the complex of streptomyces subtilisin inhibitor (SSI) and subtilisin. Structural dada, which is essential for understanding the inhibitor mechanism of SSI,was obtained
1.The methods which we had proposed, MLEV-4,4', MLEV8/8^R, RACO,and R2TR,were further developed to be used as practical tools.
2.A theory, which is vital to manipulate internal Hamiltonians by modulating amplitudes, phases, and frequencies of a radio-frequency field applied to spins, was developed. This method was applied successfully to recover anisotropic chemical shift or ^<13>C-^<15>N dipolar powder patterns under fast magic angle spinning.
3.Lattice vibration effects on distances measured by X-ray diffraction and solid-state NMR were theoretically examined.
4.The complex of streptomyces subtilisin inhibitor (SSI) and subtilisin was prepared, where the carbon and the nitrogen at the scissile bond were selectively replaced by their isotopes (^<13>C and ^<15>N).
5.The ^<13>C-^<15>N distance of the labeled SSI-subtilisin complex sample was precisely determinded, and an accelerate structure of the labeled region of the complex was deduced.
6.The structure determined as above and that obtained by X-ray diffraction were compared to discuss the inhibitor mechanism.
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