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

基因组研究后的一个主题是确定蛋白质3D结构。这项研究的主要技术是X射线晶体学。 X射线晶体学在分析晶体方面取得了巨大进展，因为同步辐射已被利用。鉴于，制备高质量单晶的技术仍然日益进步。我们证明了通过XE-LAMP照射几十秒钟的XE-LAMP辐照，鸡蛋白溶菌酶成核的光化学诱导的成核。我们已经证明了核增强机制。^<1）>我们在这里报告，1）改进实验程序，具有良好的可重现性，2）更适合光诱导的核核的结晶食谱的投资； 3）3）除母鸡鸡蛋溶菌酶外，光诱导的核蛋白。 [结果] 1）改进实验程序。蛋白质结晶实验通常是通过蒸气扩散进行的，例如悬挂掉或坐下方法。 …更多，我们首先试图照射光目录以在玻璃板上过饱和溶液。但是，观察到核增强功能，可重复性差。这是由于液滴的蒸气液界面蒸发而造成的，这引起了不均匀的成核。为了避免这种不确定性，我们改进了图中所示的实验程序。结果，我们获得了良好的可重复性，即在没有光照射的情况下将成核不会发生在溶液中。另一方面，成核是通过辐照溶液进行的。2）研究光诱导成核的结晶食谱。我们研究了适合光诱导成核的结晶食谱。聚乙烯甘油4000（PEG4000）比在同一过饱和的情况下含有PEG4000的情况更有效地增强了光诱导的成核。3）Thaumatin的结晶。作为另一种蛋白质，除了鸡蛋溶菌酶外，还进行了丁马廷的结晶。确定溶解度曲线。即使在不饱和溶液中，成核增强也暂时也发生。这表明丁胺蛋白之间的分子相互作用因辐照而剧烈变化。 [结论]改进了实验程序。对几种商业上可获得的蛋白质进行了光诱导的核实验。我们还计划开发工业蛋白质结晶系统。较少的

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