Charge Detection Mass Spectrometry Identifies Preferred Non-Icosahedral Polymorphs in the Self-Assembly of Woodchuck Hepatitis Virus Capsids.
Charge Detection Mass Spectrometry Identifies Preferred Non-Icosahedral Polymorphs in the Self-Assembly of Woodchuck Hepatitis Virus Capsids.
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DOI：
10.1016/j.jmb.2015.06.019
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发表时间：
2016-01-29
影响因子：
5.6
通讯作者：
Jarrold MF
中科院分区：
文献类型：
--
作者：
Pierson EE;Keifer DZ;Kukreja AA;Wang JC;Zlotnick A;Jarrold MF
土拨鼠肝炎病毒（WHV）在体外容易发生异常组装，并且可以形成广泛分布的超大颗粒。表征异常装配产品具有挑战性，因为它们既庞大又异构。在这项工作中，电荷检测质谱 (CDMS) 用于测量 WHV 组装产品的分布。 CDMS 是一种单粒子技术，通过同时测量每个离子的电荷和 m/z 比来确定单个离子的质量。在相对积极的组装促进条件下，大约一半的 WHV 组装产物是由 120 个二聚体组成的 T=4 衣壳，而另一半是广泛分布的较大物种，延伸到超过 210 个二聚体。在 132 个二聚体和 150 个二聚体附近有明显的峰。在某种程度上，150 个二聚体复合物可归因于通过添加六聚体环沿其五重轴延长 T=4 衣壳。然而，大多数其他特征无法用现有的六聚体缺陷模型来解释。冷冻电子显微镜提供了细长衣壳的证据。然而，图像分析表明，其中许多并不是封闭的，而是具有“螺旋状”形态。 CDMS 数据表明，超大衣壳倾向于通过添加 3 或 4 个二聚体（可能是通过完成六聚体顶点）来生长。
Woodchuck hepatitis virus (WHV) is prone to aberrant assembly in vitro, and can form a broad distribution of oversized particles. Characterizing aberrant assembly products is challenging because they are both large and heterogeneous. In this work, charge detection mass spectrometry (CDMS) is used to measure the distribution of WHV assembly products. CDMS is a single particle technique where the masses of individual ions are determined from simultaneous measurement of each ion’s charge and m/z ratio. Under relatively aggressive assembly-promoting conditions, roughly half of the WHV assembly products are T=4 capsids composed of exactly 120 dimers while the other half are a broad distribution of larger species that extends to beyond 210 dimers. There are prominent peaks at around 132 dimers and at 150 dimers. In part, the 150 dimer complex can be attributed to elongating a T=4 capsid along its five-fold axis by adding a ring of hexamers. However, most of the other features cannot be explained by existing models for hexameric defects. Cryo-electron microscopy provides evidence of elongated capsids. However, image analysis reveals that many of them are not closed, but have “spiral-like” morphologies. The CDMS data indicates that oversized capsids have a preference for growth by addition of 3 or 4 dimers, probably by completion of hexameric vertices.
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DOI：
10.1073/pnas.0400444101
发表时间：
2004-04-20
影响因子：
11.1
作者：
Fokine, A;Chipman, PR;Rossmann, MG
通讯作者：
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影响因子：
3.7
作者：
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影响因子：
3.7
作者：
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通讯作者：
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影响因子：
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通讯作者：
MELLEMA, JE
影响因子：
3.4
作者：
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通讯作者：
Reguera, David