Millisecond Order Dynamics of Protein Crystal Structure Using Syncrotron Radiation.
Millisecond Order Dynamics of Protein Crystal Structure Using Syncrotron Radiation.
批准号:
05244104
负责人:
SAKABE Noriyoshi
金额:
$26.56万
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research on Priority Areas
财政年份:
1993
资助国家:
日本
项目状态:
已结题
起止时间:
1993 至 1997
中文摘要
与x射线晶体结构分析仅阐明结构的静态方面的普遍观点相反,本项目旨在以毫秒级的时间分辨率阐明蛋白质反应过程中的时间依赖性结构,从而通过劳厄方法从三维结构变化的角度理解这种反应过程。从1993年到1996年,我们对这个困难的研究进行了4年的广泛挑战。在这个研究项目的最后,我们可以总结我们的成功在以下五个方面;(1)构建了基于大分子定向时间分辨劳埃相机的数据采集系统,该系统也可用于PF中弯曲磁束线BL-18B的单色x射线Weissenberg相机;(2)基于CCP4的劳埃套件开发数据处理软件;(3)建立了各种辅助技术,包括各种软件的应用方法,以及结晶技术,特别是膜蛋白;(4)通过时间分辨劳法接近阐明反应过程;(5)对许多蛋白质的反应过程进行结构分析,这是非常重要的研究。这些结构分析中最突出的结果之一是tsukahara教授对牛肉细胞色素c氧化酶的分析。最后,应用上述仪器和技术对血红素蛋白、RNA降解酶和核苷酸结合蛋白等几种蛋白质系统的反应和调控机制进行了研究。最突出的工作是阐明了谷氨酸合成的时间依赖结构。这些结果报告在最终项目报告中,由s Ebashi教授领导的咨询小组出版了5年的出版物和会议记录,其中展示了来自不同科学领域的专家如何开发一个新领域,例如物理学,晶体学,有机合成,物理化学家,MNR专家,生物化学家,理论物理学家和光谱学家,以及出版物和研讨会的简要内容。少
英文摘要
Contrary to the prevailing idea that x-ray crystal structure analyzes elucidate only the static aspects of structures, the present project aims at elucidating the time dependent structures of proteins in their processes of reaction at a millisecond order time-resolution and thus understanding such processes of reaction in terms of three-dimensional structur change by Laue method. We have challenged this difficult study for 4 years from 1993 to 1996, extensively. At the end of this research project, we can summarize our success in following five items ; (1) construction of a data collection system based on a macromolecule-oriented time-resolved Laue camera which we can also use for Weissenberg camera with monochromatic X-rays at the bending magnet beam line BL-18B in the PF,(2) development of data processing software based on the Laue suites of CCP4, (3) establishment of various supporting techniques including application tequnique of various softwares for X-ray crystal structure analys … More is and crystallization techniques especially membrane protein, (4) comming close to elucidate the reaction process by the time-resolved Laue method, (5) Structure analyzes of many proteins reaction proccess of which are very important to study. One of the most prominent result of these structure analyzes is that of cytocrome c oxydase from beef by prof.Tsukahara.Finally above instrument and techniques were applied to study the several kinds of protein systems including haem proteins, RNA degrading enzymes, and nucleotide-binding proteins in terms of their reaction and control mechanism. The most prominent work is the elucidating time dependent structure of glutatione syntheses by Prof.Oda. These results are reported in the final project report with publications and minute books publised by the advisery group leaded by Prof.S Ebashi for 5 years where it shows how a new field has been developed by the corporation of specialists from various scientific field, such as physics, crystallography, organic synthesis, physical chemist, MNR specialist, biochamist, theoretical physicist and spectroscopists, and a brief content of publications and symposiums. Less
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Sakabe K., K.Sasaki, N.Watanabe, M.Suzuki, Z.G.Wang, J.Miyahara & N.Sakabe: "Large-Format Imaging Plate and Weissenberg Camera for Accurate Protein Crystallographic Data Collection Using Synchrotron Radiation" J.Synchrotron Rad.4. 136-146 (1997)
Sakabe K.、K.Sasaki、N.Watanabe、M.Suzuki、Z.G.Wang、J.Miyahara
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