A Rapid Analysis System of Protein Crystal Structures

蛋白质晶体结构快速分析系统

• 批准号: 01880019
• 负责人: NAKAMURA Kazuo
• 金额: $ 2.94万
• 依托单位: Nagaoka University of Technology (1990)The University of Tokyo (1989)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B).
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01880019/
• 关键词: Molecular graphics; Protein; Three-dimensional structure; X-ray analysis; Refinement

1. After a personal IRIS graphics workstation was introduced into our laboratory, we checked its graphic and arithmetic performances at first. In result, we found that it has enough performance to realize the object of this project. Then we adapted the TOM program, a graphics fitting program for macromolecules originally coded by Cambillau (1987), to the Personal IRIS workstation. 2. Using a mainframe computer, we optimized thoroughly the stereochemically restrained least-squares refinement program PROLSQ (Hendrickson and Konnert, 1980) modified by us so that the execution time might become as short as possible. After the optimization we adapted the PROLSQ program to the Personal IRIS workstation, and improved the interface between the PROLSQ and TOM programs so that crystallographers might refine protein structures easily and quickly. 3. In order to see if this refinement system works well, we refined the crystal structure of the complex between microbial ribonuclease Ms and its inhibitor, 3'-guanylic acid, using the system. Its coarse atomic coordinates had been obtained by molecular replacement methods. The refinement system worked quite efficiently. It is the proof of the efficiency of the refinement system that for the past only one year we have finished refining seven kinds of protein structures, (1) RNase Ms*3'-GMP complex (2) Subtilisin-SSI complex (3) RNase Ms*2'-GMP complex (4) RNase Ms*GfpC complex (5) Trypsin-SSI complex (6) Interferon-beta and (7) RNase Rh, using this refinement system.

1、实验室引进个人IRIS图形工作站后，我们首先对其图形和运算性能进行了测试。结果，我们发现它有足够的性能来实现这个项目的目标。然后我们将TOM程序（最初由Cambillau（1987）编码的大分子图形拟合程序）改编到Personal IRIS工作站。 2.利用大型计算机对我们修改的立体化学约束最小二乘精化程序PROLSQ (Hendrickson and Konnert, 1980)进行了彻底优化，使执行时间尽可能缩短。优化后，我们将PROLSQ程序适配到Personal IRIS工作站中，并改进了PROLSQ和TOM程序之间的接口，以便晶体学家可以轻松快速地精炼蛋白质结构。 3. 为了验证该精制系统是否有效，我们利用该系统对微生物核糖核酸酶Ms与其抑制剂3'-鸟苷酸之间的复合物的晶体结构进行了精制。通过分子置换方法获得了其粗略的原子坐标。精炼系统运转得相当高效。仅用一年时间我们就完成了七种蛋白质结构的精制，（1）RNase Ms*3'-GMP复合物（2）Subtilisin-SSI复合物（3）RNase，这是精制系统效率的证明Ms*2'-GMP 复合物 (4) RNase Ms*GfpC 复合物 (5) 胰蛋白酶-SSI 复合物 (6) 干扰素-β 和 (7) RNase Rh，使用此精制系统。

项目成果

Yasuo Takeuchi: "Refined Crystal Strueture of the Complex of Subtilisin BPN' and Streptomyces Subtilisin Inhibitor at 1.8A^^° Resolution" J.Mol.Biol.(1991)

Yasuo Takeuchi：“枯草杆菌蛋白酶 BPN 和链霉菌枯草杆菌蛋白酶抑制剂复合物在 1.8A^^° 分辨率下的精制晶体结构”J.Mol.Biol.(1991)

中村和郎: "結晶の分子科学入門(第5章第3節)" 講談社サイエンティフク, 236 (1989)

中村一夫：“晶体分子科学导论（第 5 章第 3 节）”讲谈社科学福，236（1989）

Yasuo Takeuchi: "Refined Crystal Structure of the Complex of Subtilisim BPN' and Streptomyces Subtilisin Inhibitor at 1.8A Rosolution" J. Mol. Biol.(1991)

Yasuo Takeuchi：“1.8A Rosolution 下枯草杆菌 BPN 和链霉菌枯草杆菌蛋白酶抑制剂复合物的精制晶体结构”J. Mol。

中村 和郎: "新しい実験化学講座 10「回折」4章5節「モデル作成」" 丸善(株), (1992)

中村和夫：《新实验化学教程 10“衍射”第 4 章第 5 节“模型创建”》丸善株式会社，（1992 年）

Hiroyuki Kurihara: "Crystallization of a New Class of Microbial Ribonuclease from Rhizopus Niveus" J.Mol.Biol.206. 791-792 (1989)

Hiroyuki Kurihara：“来自尼维根霉的一类新微生物核糖核酸酶的结晶”J.Mol.Biol.206。