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口蹄疫病毒的结晶及初步X射线衍射分析

Crystallization and preliminary X-ray diffraction analysis of foot-and-mouth disease virus.

作者信息

Fox G, Stuart D, Acharya K R, Fry E, Rowlands D, Brown F

机构信息

Wellcome Biotechnology Ltd., Pirbright, Surrey, England.

出版信息

J Mol Biol. 1987 Aug 5;196(3):591-7. doi: 10.1016/0022-2836(87)90035-0.

Abstract

Foot-and-mouth disease virus has been crystallized with the objectives of (1) determining the composition and conformation of the major immunogenic site(s) and (2) comparing its structure with those of the related polio, rhino and Mengo viruses, representing the other three genera of the picornaviruses. Most of the work has been done with virus strain O1BFS 1860, which crystallized as small rhombic dodecahedra of maximum dimension 0.3 mm. Virus recovered from crystals was infectious, and was indistinguishable from native virus both in protein composition and buoyant density. The stability of the crystals in the X-ray beam was comparable with that of other picornavirus crystals and they diffracted to a resolution of better than 2.3 A. Initial analysis of the X-ray diffraction data shows the virus to be positioned on a point of 23 symmetry in a close-packed array so that examples of all the icosahedral symmetry elements, except the 5-fold axes, are expressed crystallographically. The cell dimensions are a = b = c = 345 A, alpha = beta = gamma = 90 degrees, with a space group of I23. The diameter of the virus particle is 300 A. Despite the small size of the crystals, diffraction data have been collected to a reasonable resolution using a synchrotron source. Phasing of the diffraction data will be attempted using the methods of molecular replacement.

摘要

口蹄疫病毒已被结晶,目的如下:(1)确定主要免疫原性位点的组成和构象;(2)将其结构与相关的脊髓灰质炎病毒、鼻病毒和门戈病毒的结构进行比较,这三种病毒代表了小核糖核酸病毒科的其他三个属。大部分工作是用O1BFS 1860病毒株完成的,该病毒结晶为最大尺寸为0.3毫米的小菱形十二面体。从晶体中回收的病毒具有传染性,在蛋白质组成和浮力密度方面与天然病毒没有区别。晶体在X射线束中的稳定性与其他小核糖核酸病毒晶体相当,并且它们的衍射分辨率优于2.3埃。对X射线衍射数据的初步分析表明,病毒位于紧密堆积阵列中的23对称点上,因此除了5次轴外,所有二十面体对称元素的实例都在晶体学上得到了表达。晶胞尺寸为a = b = c = 345埃,α = β = γ = 90度,空间群为I23。病毒粒子的直径为300埃。尽管晶体尺寸较小,但已使用同步辐射源收集到了分辨率合理的衍射数据。将尝试使用分子置换方法对衍射数据进行相位分析。

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Structure determination of Mengo virus.门戈病毒的结构测定
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