Establishment of X-Ray Structure Analysis System for Protein Crystals by Multiwavelength Anomalous Diffraction Methods.

多波长反常衍射方法建立蛋白质晶体X射线结构分析系统。

基本信息

批准号：
02402051
负责人：
SATOW Yoshinori
金额：
$ 15.68万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

The objective of this. research project is to establish a system for direct and rapid structure determination of protein crystals by use of tunable and intense synchrotron X-radiation.First, a dead time correction method for scintillation detectors and automatic optimization procedures for alignment of the X-ray optical components were newly developed for the diffraction intensity measurement system, which consists of the double-crystal monochromator, focussing mirror, and horizontal-type four-circle diffractometer, installed at Photon Factory, National Laboratory for High Energy Physics. The measurement system improved so as to accommodate these procedures is applicable to four-circle diffractometry as well as to oscillation methods, and provides excellent-quality diffraction data at any practical wavelengths. The system has been in open use to various synchrotron experiments. Next, a kappa-axis diffractometer-with a sealed X-ray tube was equipped with a portable crystal-cooling unit … More which decreases radiation damages on protein crystals. Finally, computer programs for processing diffraction data measured at several wavelengths as well as for direct phasing of crystal structure factors were developed. The programs combined with structure refinement, graphics, and other programs were built into a complete system for multiwavelength anomalous diffraction analyses.Crystal Structure of an Fv fragment from mouse anti-dansyllysine immunoglobulin IgG2a has been determined at 2.5 * resolution. The structure clearly elucidates its complementarity determining regions. Structure refinements of the native crystal and a dansyllysine complex crystal are in progress at hislier resolution. Furthermore, crystal structures of ferous spincrossover complexes as well as those of aspartaine IA and IB forms liave been solved in the course of performance tests of the measurement and proprain systems.The systems established in this research project are widely applicable to crystal structure studies of proteins and others. Less

目的。研究项目将通过使用可调节和强烈的同步器X辐射来建立一个系统，以直接和快速的结构确定蛋白质晶体。水平型四环衍射仪，安装在高能物理国家实验室的Photon Factory。改进了测量系统以适应这些过程，适用于四环衍射法以及振荡方法，并在任何实际波长下提供出色的质量衍射数据。该系统已公开用于各种同步器实验。接下来，带有密封的X射线管的Kappa轴衍射仪配备了便携式晶体冷却单元……更多可减少蛋白质晶体的辐射损害。最后，开发了用于处理以几个波长以及直接相拟定晶体结构因子的处理衍射数据的计算机程序。与结构改进，图形和其他程序相结合的程序已内置在多波其多波长异常衍射分析的完整系统中。已在2.5 *分辨率下确定了小鼠抗二酰胺免疫球蛋白IgG2A的FV片段结构。该结构清楚地阐明了其完整性确定区域。在更高的分辨率下，天然晶体和丹酰胺复合物晶体的结构改进正在进行中。此外，在该研究项目中建立的系统广泛适用于蛋白质和其他晶体结构研究中，在绩效测试的过程中，在绩效测试的过程中求解了稀疏的旋转杂交配合物以及Aspartine IA和IB形式的晶体结构。较少的