Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo.
Averaging of viral envelope glycoprotein spikes from electron cryotomography reconstructions using Jsubtomo.
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DOI：
10.3791/51714
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发表时间：
2014-10-21
期刊：
影响因子：
--
通讯作者：
Bowden TA
中科院分区：
文献类型：
--
作者：
Huiskonen JT;Parsy ML;Li S;Bitto D;Renner M;Bowden TA
包膜病毒利用其表面的膜糖蛋白介导进入宿主细胞。这些糖蛋白“刺突”的三维结构分析通常在技术上具有挑战性，但对于理解病毒发病机制和药物设计很重要。在这里，提出了一种通过电子冷冻断层扫描数据的计算平均来确定病毒尖峰结构的方案。电子冷冻断层扫描是电子显微镜中的一种技术，用于获得近天然冷冻水合状态的膜病毒等多形生物样本的三维断层体积重建或断层图像。这些断层图像揭示了三维感兴趣的结构，尽管分辨率很低。子体积或子断层图像的计算平均值对于获得重复结构基序的更高分辨率细节是必要的，例如病毒糖蛋白尖峰。一个详细的计算方法对齐和平均使用Jsubtomo软件包的子断层图像的概述。这种方法使得病毒糖蛋白刺突的结构的可视化能够达到20-40 μ m范围内的分辨率，并且能够研究病毒体膜上的高阶刺突-刺突相互作用。典型的结果是布尼亚韦拉病毒，包膜病毒从家庭布尼亚病毒科。该家族是一组结构多样的病原体，对人类和动物健康构成威胁。
Enveloped viruses utilize membrane glycoproteins on their surface to mediate entry into host cells. Three-dimensional structural analysis of these glycoprotein ‘spikes’ is often technically challenging but important for understanding viral pathogenesis and in drug design. Here, a protocol is presented for viral spike structure determination through computational averaging of electron cryo-tomography data. Electron cryo-tomography is a technique in electron microscopy used to derive three-dimensional tomographic volume reconstructions, or tomograms, of pleomorphic biological specimens such as membrane viruses in a near-native, frozen-hydrated state. These tomograms reveal structures of interest in three dimensions, albeit at low resolution. Computational averaging of sub-volumes, or sub-tomograms, is necessary to obtain higher resolution detail of repeating structural motifs, such as viral glycoprotein spikes. A detailed computational approach for aligning and averaging sub-tomograms using the Jsubtomo software package is outlined. This approach enables visualization of the structure of viral glycoprotein spikes to a resolution in the range of 20-40 Å and study of the study of higher order spike-to-spike interactions on the virion membrane. Typical results are presented for Bunyamwera virus, an enveloped virus from the family Bunyaviridae. This family is a structurally diverse group of pathogens posing a threat to human and animal health.
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影响因子：
64.8
作者：
Liu, Jun;Bartesaghi, Alberto;Borgnia, Mario J.;Sapiro, Guillermo;Subramaniam, Sriram
通讯作者：
Subramaniam, Sriram
DOI：
10.1083/jcb.201104040
发表时间：
2011-11-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Karotki L;Huiskonen JT;Stefan CJ;Ziółkowska NE;Roth R;Surma MA;Krogan NJ;Emr SD;Heuser J;Grünewald K;Walther TC
通讯作者：
Walther TC
影响因子：
3
作者：
Kremer, JR;Mastronarde, DN;McIntosh, JR
通讯作者：
McIntosh, JR
影响因子：
5.4
作者：
Huiskonen, Juha T.;Hepojoki, Jussi;Gruenewald, Kay
通讯作者：
Gruenewald, Kay
影响因子：
6.8
作者：
Briggs, John A. G.
通讯作者：
Briggs, John A. G.