Investigation of light-induced nucleation mechanism of proteins

蛋白质光诱导成核机制的研究

基本信息

批准号：
16550005
负责人：
OKUTSU Tetsuo
金额：
$ 2.37万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

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

后基因组研究的一个主题是确定蛋白质的 3D 结构，自从使用同步辐射制备高质量单晶以来，X 射线晶体学在分析晶体方面取得了巨大进展。我们已经证明，通过氙灯照射几十秒，光化学诱导鸡蛋清溶菌酶成核。 ^<1)> 我们在此报告，1) 实验程序的改进，具有良好的重现性，2) 研究更适合光诱导成核的结晶配方，3) 蛋白质的光诱导成核，除了【结果】1）改进了实验流程。蛋白质结晶实验一般采用悬滴法或坐滴法等蒸气扩散法进行。首先，我们尝试用光照射玻璃板上的过饱和溶液，尽管观察到成核增强，但再现性很差，这是由于液滴的蒸气-溶液界面蒸发导致不均匀成核造成的。为了避免这种不确定性，我们改进了如图所示的实验程序，结果，我们获得了良好的重现性，即在没有光的情况下，溶液中不会发生成核。 2) 光诱导成核的结晶配方研究我们研究了适合光诱导成核的结晶配方，比光诱导成核更有效。相同过饱和度下不含有PEG4000。3)奇异果甜蛋白的结晶作为除此之外的另一种蛋白质。鸡蛋溶菌酶中，发现了索马甜的溶解度曲线，即使在不饱和溶液中也发生了暂时的成核增强，这表明索马甜之间的分子相互作用通过照射而发生了有吸引力的变化。我们还计划开发工业化蛋白质结晶系统。

项目成果

期刊论文数量（64）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Light-induced nucleation of metastable hen-egg white lysozyme solutions

亚稳态鸡蛋清溶菌酶溶液的光诱导成核

DOI：
发表时间：
2005
期刊：
Crystal Growth and Design 5
影响因子：
0
作者：
T.Okutsu;K.Furuta;M.Terao;H.Hiratsuka;A.Yamano;N.Ferte;S.Veesler
通讯作者：
S.Veesler

Photophysical and Photochemical Processes of 9,10-Dihydro-9-silaphenanthrene Derivatives : Photochemical Formation and Electronic Structure of 9-Silaphenanthrene

9,10-二氢-9-硅菲衍生物的光物理和光化学过程：9-硅菲的光化学形成和电子结构

DOI：
发表时间：
2006
期刊：
J.Phys.Chem.A 110
影响因子：
0
作者：
Hiroshi Hiratsuka;Hiroaki Horiuchi Yasunari Frukawa;Hikaru Watanabe;Akiko Ishihara;Tetsuo Okutsu;Seiji Tobita;Tetsutaro Yoshinaga;Akihiro Shinohara;Norihiro Tokitoh;Makoto Oba;Kozaburo Nishiyama
通讯作者：
Kozaburo Nishiyama

光科学研究の最前線

光学科学研究的前沿