1988 Fiscal Year Final Research Report Summary
Crystallization of Proteins for X-ray Analysis and the Accurate Intensity Data Measurement
| Project/Area Number |
61304062
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
結晶学
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| Research Institution | Tokyo Institute of Technology (1988)
Osaka University (1986-1987) |
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Principal Investigator |
TANAKA Nobuo Tokyo Institute of Technology, Faculty of Science, 理学部, 教授 (50032024)
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| Co-Investigator(Kenkyū-buntansha) |
YASUOKA Noritake Himezi Institute of Technology, Institute of the Engineering, 工学基礎研究所, 教授 (40029054)
UEKI Tatsuo Osaka University, Faculty of Engineering Science, 基礎工学部, 助教授 (30029954)
KAI Yatushi Osaka University, Faculty of Engineering, 工学部, 助教授 (40029236)
AIBARA Shigeo Kyoto University, The Research Institute of Food Science, 食糧科学研究所, 助教授 (20027197)
TAKENAKA Akio Tokyo Institute of Technology, Faculty of Science, 理学部, 助手 (80016146)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Crystallization of Proteins / X-ray Crystal Structure Analysis / 精密回析データの測定 |
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| Research Abstract |
We have succeeded to produce big crystals from many kinds of proteins, which were suitable for X-ray crystal structure analysis, myoglobins reconstituted by four kinds of synthetic hemes were crystallized by both dialysis and vapor diffusion methods. In case of the vapor diffusion method, the film on the surface obstructs to grow the cycstals. The intensity of reflection was measured by the diffractometer and the Weissenberg camera of the photon factory at Tsukuba. The structures refined with each data were not significantly different with each other, which indicates that any method will go for the structure analysis. The structure was solved by the molecular replacement method. In spite of different hemes, the apo proteins had the same structure as that of native sperm myoglobin. The doughnuts-like electron density distribution was obtained for the porphine reconstituted molecule. Probably, the home rotates along the ligated axis, as expected from the NMR spectroscopy. The other proteins studied in this projects are: 3-ispropylmalate dehydrogenase of Thermus thermophilus, lipoamide dehydrogenase, Cu,Zn and Mn-superoxide dismutases, protease-Inhibitor complex, protease modified by site-directed mutagenesis, hydrogenase, ribonuclease Rh so on.
The IP system was developed with the software to convert the measured data to the structure amplitude of each reflection. This system proved to work well by the data measurement of lysozyme. The direct phase determination by the multi-wave measurement of the synchrotron X ray was also developed. These new technigues will be utilized by many researchers.
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