Development of Rapid and Accurate Methods for X-ray Crystallography to Study Crystal Structures and Functions of Proteins.

开发快速准确的 X 射线晶体学方法来研究蛋白质的晶体结构和功能。

基本信息

批准号：
06305006
负责人：
SATOW Yoshinori
金额：
$ 5.95万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

This research project aimed at the development of rapid and accurate methods for macromolecular X-ray crystallography, through collaborative studies of three-dimensional structures and functions of proteins. The project was carried out by 15 investigators.The proteins studied in the project were, genetically engineered proteins with amino-acid substitutions, proteolytic and glycolytic enzymes as well as their complexes with inhibitors, proteins that recognize nucleic acids, antibodies and their complexes with haptens, membrane proteins, viruses, and glycoproteins. The methods and techniques developed were, for engineering and preparation of protein specimens, for crystallization of proteins, for determination of very large structural units, for diffraction data collection, for utilization of anomalous scattering effects, for automated molecular-replacement solutions, and for utilization of other structural information.Structure determinations of a membrane protein of bovine cytochrome c oxidase and a glycoprotein of human renal dipeptidase are typical examples of the successful results. The former protein was crystallized and solved with the techniques developed. The later were prepared with genetic engineering and enzymatic deglycosylation techniques. The structures of chimeric enzymes, proteinases of a trypsin family, and unliganded and hapten-liganded forms of Fab and Fv fragments of antibodies were also determined. The developments of hardware and software systems with rapid and accurate data collection capabilities, of techniques for locating anomalous scatterers and for MAD analyzes, and of automated molecular-replacement procedures were carried out.In each year, a meeting for exchanging information about research activities of investigators was held. A report on research activities, progresses, and achievements was published at the end of each year. The report for the final year was circulated to researchers in related fields.

该研究项目旨在通过蛋白质三维结构和功能的合作研究，开发快速、准确的大分子 X 射线晶体学方法。该项目由15名研究人员进行。该项目研究的蛋白质有氨基酸取代的基因工程蛋白质、蛋白水解酶和糖酵解酶及其与抑制剂的复合物、识别核酸的蛋白质、抗体及其与半抗原的复合物、膜蛋白、病毒和糖蛋白。开发的方法和技术用于蛋白质样本的工程和制备、蛋白质的结晶、非常大的结构单元的测定、衍射数据收集、异常散射效应的利用、自动分子替换解决方案以及利用牛细胞色素c氧化酶的膜蛋白和人肾二肽酶的糖蛋白的结构测定是成功结果的典型例子。以前的蛋白质通过所开发的技术被结晶并溶解。后者是通过基因工程和酶促去糖基化技术制备的。还确定了嵌合酶、胰蛋白酶家族的蛋白酶以及抗体的 Fab 和 Fv 片段的未配体和半抗原配体形式的结构。开发了具有快速、准确数据收集能力的硬件和软件系统、定位异常散射体和 MAD 分析的技术以及自动分子替换程序。每年都会召开一次会议，交换有关研究活动的信息调查人员被拘留。每年年底都会发布一份关于研究活动、进展和成果的报告。最后一年的报告已分发给相关领域的研究人员。