Purification, crystallization and preliminary X-ray analysis of nonstructural protein 2 (nsp2) from avian infectious bronchitis virus.

Kun Yu, Zhenhua Ming, Yuanyuan Li, Cheng Chen, Zehua Bao, Zhilin Ren, Bofeng Liu, Wei Tao, Zihe Rao, Zhiyong Lou
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引用次数: 11

Abstract

Avian infectious bronchitis virus (IBV) is a member of the group III coronaviruses, which differ from the other groups of coronaviruses in that they do not encode the essential pathogenic factor nonstructural protein 1 (nsp1) and instead start with nsp2. IBV nsp2 is one of the first replicase proteins to be translated and processed in the viral life cycle; however, it has an entirely unknown function. In order to better understand the structural details and functional mechanism of IBV nsp2, the recombinant protein was cloned, overexpressed in Escherichia coli, purified and crystallized. The crystals diffracted to 2.8 Å resolution and belonged to space group P2(1), with unit-cell parameters a = 57.0, b = 192.3, c = 105.7 Å, β = 90.8°. Two molecules were found in the asymmetric unit; the Matthews coefficient was 3.9 Å(3) Da(-1), corresponding to a solvent content of 68.2%.

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禽传染性支气管炎病毒非结构蛋白2 (nsp2)的纯化、结晶及x射线初步分析。
禽传染性支气管炎病毒(IBV)是III族冠状病毒的一员,与其他冠状病毒群不同的是,它们不编码必需致病因子非结构蛋白1 (nsp1),而是从nsp2开始编码。IBV nsp2是最早在病毒生命周期中被翻译和加工的复制酶蛋白之一;然而,它有一个完全未知的功能。为了更好地了解IBV nsp2的结构细节和功能机制,对重组蛋白进行了克隆、在大肠杆菌中过表达、纯化和结晶。晶体衍射分辨率为2.8 Å,属于P2(1)空间群,晶胞参数a = 57.0, b = 192.3, c = 105.7 Å, β = 90.8°。在不对称单元中发现了两个分子;马修斯系数为3.9 Å(3) Da(-1),溶剂含量为68.2%。
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期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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