Determining a molecular pathway for formation of a T=3 capsid using mass spectrometry

使用质谱法确定 T=3 衣壳形成的分子途径

基本信息

批准号：
BB/E008070/1
负责人：
Peter Stockley
金额：
$ 49.53万
依托单位：
University of Leeds
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2007
资助国家：
英国
起止时间：
2007 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FE008070%2F1
关键词：
Determining molecular pathway formation T=3

项目摘要

Viruses remain major human, animal and plant pathogens. The most successful current anti-viral therapies rely on traditional vaccination, a technique first identified in the West over a century ago, and unfortunately not simply applicable to many viruses. More modern treatments are directed at inhibiting specialised enzymes within viruses with small molecular weight compounds, anti-viral drugs, such as those used to treat AIDS. Unfortunately, such enzymes usually share many features with host molecules and as a result most anti-viral drugs show unwanted side-effects. In fact viruses offer a unique molecular target that has so far not been exploited therapeutically, namely their assembly pathways. Simple viruses that contain only a nucleic acid enclosed in a protein shell form those shells as symmetrical aggregates from very few different types of subunits. The overall particle has a spherical cross-section and looks somewhat like a soccer ball. Similar structures form within more complex membrane-containing viruses as well. Despite a wealth of information about viral structures it has not yet been possible to map out experimentally an assembly pathway from protein subunit to shell in any virus. In part this is because it has traditionally been technically difficult to study this aspect of viral life-cycles. One key feature of assembly intermediates is their different molecular masses. In our model virus, a bacteriophage that normally infects a laboratory bacterium, but whose architecture mimics that of viruses that infect humans, we have recently discovered how to slow the assembly process in a test tube and measure the masses of all the intermediates formed in the pathwayto the final spherical shell using a modern mass spectrometer. This assay now allows us to take the most detailed look at the assembly pathway, examine how both the phage coat protein and RNA contribute to the assembly process, and hopefully identify generic principles that could reveal novel anti-viral drug targets.

病毒仍然是人类、动物和植物的主要病原体。目前最成功的抗病毒疗法依赖于传统的疫苗接种，这种技术在一个多世纪前首次在西方发现，不幸的是并不简单地适用于许多病毒。更现代的治疗方法旨在用小分子量化合物、抗病毒药物（例如用于治疗艾滋病的药物）抑制病毒内的特殊酶。不幸的是，此类酶通常与宿主分子具有许多共同特征，因此大多数抗病毒药物都会表现出不良副作用。事实上，病毒提供了迄今为止尚未在治疗上利用的独特分子靶点，即它们的组装途径。仅包含封装在蛋白质壳中的核酸的简单病毒将这些壳形成为由极少数不同类型的亚基对称聚集体。整个颗粒具有球形横截面，看起来有点像足球。类似的结构也形成在更复杂的含膜病毒中。尽管有大量关于病毒结构的信息，但尚不可能通过实验绘制出任何病毒从蛋白质亚基到外壳的组装途径。部分原因是传统上研究病毒生命周期的这一方面在技术上很困难。组装中间体的一个关键特征是它们不同的分子量。在我们的模型病毒（一种通常感染实验室细菌的噬菌体，但其结构模仿感染人类的病毒的结构）中，我们最近发现了如何减慢试管中的组装过程并测量在试管中形成的所有中间体的质量。使用现代质谱仪获得最终球壳的途径。现在，这种测定方法使我们能够最详细地观察组装途径，检查噬菌体外壳蛋白和 RNA 对组装过程的贡献，并希望确定可以揭示新型抗病毒药物靶点的通用原理。