X-Ray Crystallographic Analysis of Spherical Plant Viruses

球形植物病毒的 X 射线晶体学分析

• 批准号: 61580049
• 负责人: FUKUYAMA Keiichi
• 金额: $ 1.6万
• 依托单位: Faculty of Science, Osaka University (1987)Tottori University (1986)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61580049/
• 关键词: Spherical Virus; Tobacco Necrosis Virus; X-Ray Analysis; 結晶構造; タバコネクローシスウイルス; 結晶化; X線結晶解析

Tobacco necrosis virus (TNV) is a spherical plant virus consisting of 180 copies of coat protein and a single-stranded RNA. TNV was propagated in Chenopodium quinoa, and purified by differential centrifugation, gel filtration and sucrose density centrifugation. The purified TNV showed only one band at M=30,000 in SDS-polyacrylamide gel electrophoresis, and contained homogeneous particles of a diameter of approximately 28 nm in the electron microscope. TNV crystals were produced by dialyzing against 0.4 M sodium phosphate buffer with the pH adjusted to 6.0. The TNV crystals grew in the form of dodecahedron, to a maximum of 1 mm in diameter. The diffraction patterns of the crystals have shown that the crystal belongs to cubic space group p4_232 with a=338 <Ang>. The two particles in the unit cell are rotated 90 to each other about the principal axis of the crystal situated at (0,0,0) and (1/2,1/2,1/2). The nearest distance between the particles in the unit cell is 293 <Ang>. The crystals diffract X-ray to 2.5 <Ang> resolution and showed little radiation damage. Collection of three-dimensional X-ray data and preparation of heavy atom derivatives are in progress.

烟草坏死病毒（Tobacco necrosis virus，TNV）是一种由180个拷贝的外壳蛋白和一个单链RNA组成的球形植物病毒。TNV在藜中增殖，并通过差速离心、凝胶过滤和蔗糖密度离心纯化。纯化的TNV在SDS-聚丙烯酰胺凝胶电泳中在M= 30，000处仅显示一条带，在电子显微镜下含有直径约28 nm的均匀颗粒。TNV晶体通过用pH调节至6.0的0.4M磷酸钠缓冲液透析来制备。TNV晶体以十二面体的形式生长，最大直径为1 mm。晶体的衍射图表明，晶体属于立方空间群p4_232，a=338<Ang>。晶胞中的两个粒子围绕位于（0，0，0）和（1/2，1/2，1/2）处的晶体主轴彼此旋转90 °。单位晶胞中粒子之间的最近距离是293<Ang>。该晶体将X射线吸收到2.5<Ang>分辨率，并显示出很小的辐射损伤。收集三维X射线数据和制备重原子衍生物的工作正在进行中。

项目成果

Keiichi Fukuyama: "Crystallization and Preliminary X-ray Diffraction Studies of Tobacco Necrosis Virus" Journal of Molecular Biology. 196. 961-962 (1987)

Keiichi Fukuyama：“烟草坏死病毒的结晶和初步 X 射线衍射研究”分子生物学杂志。

Keiichi Fukuyama: Journal of Molecular Biology. 196. 961-962 (1987)

福山敬一：分子生物学杂志。

福山恵一: Journal of Molecular Biology.

福山敬一：分子生物学杂志。