Development of X-ray Solution Scattering System for Time-resolved

时间分辨X射线溶液散射系统的研制

• 批准号: 09558093
• 负责人: SATO Mamoru
• 金额: $ 4.99万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09558093/
• 关键词: X-ray Solution Scattering; Small-angle X-ray Scattering; Time-resolved Measurement; Dynamical Structure Analysis; Solution Structure of Protein

M. Sato setup a sample cell for time-resolved measurement of solution X-ray scattering by modifying a stopped flow mixing cell available commercially as that suitable for time-resolved structural analysis. He also introduced a multi-layer mirror system as a X-ray optics to focus on the incident X-rays at the position of an X-ray detector. The mirror system was quite superior to that using a totally reflected focusing mirror system, which was made use of so far, and realized the incident intensity ten times as bright as that using a totally reflected focusing mirror system. The combination of the X-ray optics with the sample cell establishes an X-ray solution scattering system quite suitable for the time-resolved structural analysis of protein in solution.M. Kataoka developed, as a system for the static structure analysis of protein in solution, the method to analyze the tertiary structure of protein at non-native states and also the heavy-atom replacement method to analyze the reaction … More -intermediate structure of protein. These methods were applied to α-lacatalbumin, bacteriorhodopsin and so on, and shown to be very effective to analyze the protein structure in solution. A new algorithm system to analyze the time-resolved structure of protein in solution was also developed. The algorithm system was then applied to the time-resolved solution scattering data of photo-reaction of purple membrane, and found out the two different structures in the reverse reaction from de-protonized intermediate of Schiff base produced by light absorption.On the other hand, Y. Amemiya developed two typical types of high sensitive CCD-based area detector for real time (automated) measurements and time-resolved measurements of solution X-ray scattering; one is the system in which X-ray image intensifiers are coupled to maximize the detective quantum efficiency for time-resolved measurements, and the other is the system in which tapered optical fibers are coupled for the reduction of the image into the CCD, which is optimized for automated measurements for protein crystallography. Software systems for these CCD-based X-ray detectors to correct image distortion and non-uniformity of response were also developed. Less

M. Sato通过修改商业上可获得的适合于时间分辨结构分析的停流混合池来设置用于溶液X射线散射的时间分辨测量的样品池。他还介绍了一种多层反射镜系统作为X射线光学系统，以聚焦X射线探测器位置处的入射X射线。该反射镜系统比迄今为止使用的全反射聚焦反射镜系统优越上级，并且实现了入射强度为使用全反射聚焦反射镜系统的入射强度的10倍的亮度。将X射线光学系统与样品池相结合，建立了一个非常适合于溶液中蛋白质时间分辨结构分析的X射线溶液散射系统。作为溶液中蛋白质的静态结构分析系统，片冈开发了分析非天然状态下蛋白质三级结构的方法，以及分析反应的重原子置换法 ...更多信息 - 蛋白质的中间结构。将这些方法应用于α-乳清蛋白、细菌视紫红质等蛋白质的结构分析，结果表明，这些方法对溶液中蛋白质的结构分析是非常有效的。本文还开发了一种新的蛋白质时间分辨结构分析算法系统。将该算法系统应用于紫膜光反应的时间分辨溶液散射数据，发现光吸收产生的Schiff碱去质子化中间体在逆反应中存在两种不同的结构。Amemiya开发了两种典型的基于高灵敏度CCD的真实的实时区域探测器溶液X射线散射的（自动）测量和时间分辨测量;一种是其中耦合X射线图像增强器以使时间分辨测量的探测量子效率最大化的系统，另一种是其中耦合锥形光纤以将图像缩小到CCD中的系统，其被优化用于蛋白质晶体学的自动测量。还开发了用于这些基于CCD的X射线探测器的软件系统，以校正图像失真和响应不均匀性。少

项目成果

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D. Hamada：“通过分析 2,2,2-三氟乙醇的影响来证明蛋白质折叠中 rete-limiting 步骤的性质”Nature Struct.Biol.. Vol. 7. 58-61 (2000)

Yu. A. Gaponov: "Structure Changes in the Sol-gel Systems of hydrated oxides"J. Synchrotron Rad. 5. 962-963 (1998)

于。

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T. Oka: "Conformational Change of Helix G in the Bcteriorhodopsin Photocycle : Investigation with Heavy Atom Laveling and X-ray Diffraction"Biophys. J.. 76巻. 1018-1023 (1999)

T. Oka：“Bcteriorhodopsin 光循环中螺旋 G 的构象变化：重原子 Laveling 和 X 射线衍射的研究”Biophys J.. 76. 1018-1023 (1999)

片岡幹雄: "中性子散乱による蛋白質動力学の研究"大阪大学低温センターだより. 102巻. 8-14 (1998)

Mikio Kataoka：“通过中子散射研究蛋白质动力学”大阪大学低温中心通讯第 102 卷 8-14 (1998)。