Investigation of light-induced nucleation mechanism of proteins
| Project/Area Number |
16550005
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical chemistry
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| Research Institution | Gunma University |
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Principal Investigator |
OKUTSU Tetsuo GUNMA UNIVERSITY, Associate Professor, 工学部, 助教授 (20261860)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATSUKA Hiroshi GUNMA UNIVERSITY, Professor, 工学部, 教授 (00016156)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | crystal Growth / photochemistry / protein / light-induced nucleation / lysozyme / thaumatin / 結晶 / 結晶核形成 / Thaumatin / タンパク質の結晶化 / 光誘起核形成 / 分子間力 / 動的光散乱 |
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| Research Abstract |
One subject for post genome research is to determine protein 3D structure. The main technique for this research is X-ray crystallography. X-ray crystallography has experienced great progress in analyzing a crystal since synchrotron radiation has been utilized. Whereas, the technique for the preparation of high quality single crystal is still progress day by day. We demonstrate photochemically-induced nucleation of hen egg-white lysozyme nucleation by Xe-lamp irradiation for few tens seconds. We have demonstrated nucleation enhancement mechanism.^<1)> We report here, 1) improvement of the experimental procedure with good reproducibility, 2) investigation of crystallization recipes that are more suitable for light-induced nucleation, and 3) light-induced nucleation of proteins except for hen egg lysozyme.
【Results】
1)Improvement of the experimental procedure. Protein crystallization experiments have been generally carried out by vapor diffusion such as hanging drop or sitting drop method.
… More
We, first, tried to irradiate light directory to supersaturated solution on a glass plate. Though, nucleation enhancement was observed, reproducibility was poor. This was result from undesired nucleation by evaporation from vapor-solution interface of he droplets, which induce inhomogeneous nucleation. To avoid such uncertainties, we improved experimental procedures illustrated in Figure. As a result, we obtained good reproducibility that nucleation did not take place in the solution without light irradiation ; on the other hand nucleation took place with irradiated solution.
2) Investigation of crystallization recipes for light-induced nucleation. We investigated crystallization recipes suitable for light-induced nucleation. Polyethylene glycol 4000 (PEG4000) enhanced more effectively light-induced nucleation than without containing PEG4000 at the same supersaturation.
3)Crystallization of thaumatin. As another protein except for hen-egg lysozyme, crystallization of thaumatin was carried out. Solubility curve was determined. Nucleation enhancement took place temporary even in undersaturated solution. This suggests that molecular interaction between thaumatin changes attractively by the irradiation.
【Conclusion】
Experimental procedure was improved. Light-induced nucleation experiments are carried out for several commercially obtainable proteins. We are also planning to develop industrial protein crystallization system. Less
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Report
(3 results)
Research Products
(32 results)
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[Journal Article] Photophysical and Photochemical Processes of 9, 10-Dihydro-9-silaphenanthrene Derivatives : Photochemical Formation and Electronic Structure of 9-Silaphenanthrene2006
Author(s)
H.Hiratsuka, H.Horiuchi, Y.Frukawa, H.Watanabe, A.Ishihara, T.Okutsu, S.Tobita, T.Yoshinaga, A.Shinohara, N.Tokitoh, M.Oba, K.Nishiyama
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Journal Title
J. Phys. Chem. A 110
Pages: 3868-3874
Description
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[Journal Article] Thermal Hysteresis and Thickness Dependence of the Molecular Orientation of Poly(di-n-hexylsilane) in the Film State2006
Author(s)
H.Horiuchi, T.Ishihara, S.Tsunoda, D.Morishita, T.Takei, T.Okutsu, T.Seko, H.Sugino, Y.Kawakami, H.Watanabe, M.Abe, S.Takigami, H.Hiratsuka
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Journal Title
J. Phys. Chem. B (In press)
Description
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[Journal Article] Photophysical and Photochemical Processes of 9,10-Dihydro-9-silaphenanthrene Derivatives : Photochemical Formation and Electronic Structure of 9-Silaphenanthrene2006
Author(s)
Hiroshi Hiratsuka, Hiroaki Horiuchi Yasunari Frukawa, Hikaru Watanabe, Akiko Ishihara, Tetsuo Okutsu, Seiji Tobita, Tetsutaro Yoshinaga, Akihiro Shinohara, Norihiro Tokitoh, Makoto Oba, Kozaburo Nishiyama
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Journal Title
J.Phys.Chem.A 110
Pages: 3868-3874
Description
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[Journal Article] Thermal Hysteresis and Thickness Dependence of the Molecular Orientation of Poly(di-n-hexylsilane) in the Film State
Author(s)
Hiroaki Horiuchi, Tomihiko Ishihara, Shin-ichi Tsunoda, Daisuke Morishita, Tomoyuki Takei, Tetsuo Okutsu, Toshihiro Seko, Hiroaki Sugino, Yasuyuki Kawakami, Hamao Watanabe, Minoru Abe, Shoji Takigami, Hiroshi Hiratsuka
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Journal Title
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[Book] 光科学研究の最前線2005
Author(s)
奥津哲夫, 平塚浩士
Total Pages
2
Publisher
強光子場科学研究懇談会
Description
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[Book] 光による結晶成長2004
Author(s)
奥津哲夫
Total Pages
3
Publisher
光化学
Description
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