CHARACTERIZING VIRUSES WITH MASS SPECTROMETRY

描述：（申请人摘要）病毒结构的研究既 科学上有趣的和医学上相关的。 我们寻求获得 病毒的信息，利用固有的速度和 通过使用我们开发的技术， 来研究病毒结构 虽然病毒可以被其他人分析， 尽管使用了光谱技术，但这些方法决不是常规或完整的。 由于病毒由多个蛋白质-蛋白质亚基组成， 蛋白水解后进行MALDI-MS将用于绘制 暴露在病毒表面，并提供有关 蛋白质动力学通过与晶体学数据的比较。 为了 为了进行这些分析，我们已经开始对一个模型系统进行研究， 鸡场病毒（FHV），其是一种小RNA二十面体动物病毒， 在晶体学上得到了很好的表征。 我们的MALDI-MS/蛋白水解 该方法已经被证明对小于10 μ g的FHV是有用的， 而晶体学数据通常需要100毫克的材料。 一旦 我们将转向那些还没有 结晶的或完全特征化的，如黄瓜花叶病毒。 病毒 自催化蛋白水解事件也将被检查，通过 鉴定蛋白质片段，我们将研究它们在 引发病毒感染。 FHV还表现出一种结构特征， 称为“口袋因子”，其类似于在人类中观察到的特征， 鼻病毒 有趣的是，晶体学研究并没有揭示 有机分子的结构，很可能是由于结构紊乱或 低占有率。 然而，“口袋因素”的存在与 与FHV和鼻病毒的病毒感染性相关， 被识别。 我们的目标是调查/表征这个因素，使用 我们以前开发的方法。 除了表征病毒 子结构，我们还设计了一个质谱仪进行质量 对整个病毒的分析，初步研究使我们能够 证明病毒的超微结构和感染性在整个过程中的保留 ESI质谱分析过程。 能够大量测量整个病毒 并获得病毒的结构信息可以为病毒学家提供一个 准确的病毒鉴定和表征手段。 我们的目标是 我们的工作是开发新的方法来了解当地和全球的病毒 结构

DESCRIPTION: (applicant's abstract) The study of virus structure is both scientifically interesting and medically relevant. We seek to gain information on viruses by taking advantage of the inherent speed and accuracy of mass spectrometry and by using the techniques we have developed to study viral structure. While viruses can be analyzed by other spectroscopic techniques, those methods are by no means routine or complete. Since viruses are composed of multiple protein-protein subunits, viral proteolysis followed by MALDI-MS will be used to map the amino acids that are exposed on the viral surface and to provide information about the protein dynamics through comparisons with crystallographic data. In order to perform these analyses we have initiated studies on a model system, the flock house virus (FHV), which is a small RNA icosahedral animal virus that is well-characterized crystallographically. Our MALDI-MS/proteolysis approach has already been shown to be useful on less than 10 ug of FHV, whereas crystallographic data typically requires 100 mg of material. Once the approach is established we will move to viruses that have not been crystallized or fully characterized such as cucumber mosaic virus. Viral autocatalytic proteolysis events will also be examined and, through identification of protein fragments, we will investigate their role in initiating viral infectivity. The FHV also exhibits a structural feature termed the "pocket factor" that is analogous to a feature observed in human rhinoviruses. Interestingly, crystallographic studies do not reveal the structure of the organic molecule, most likely due to structural disorder or low occupancy. The existence of the "pocket factor", however, is closely associated with viral infectivity in FHV and rhinoviruses and has not yet been identified. Our goal is to investigate/characterize this factor using methods that we previously developed. In addition to characterizing viral substructure, we are also designing a mass spectrometer to perform mass analysis on whole viruses, preliminary research has allowed us to demonstrate the retention of virus ultrastructure and infectivity throughout the ESI mass analysis process. The ability to mass measure whole viruses and gain structural information on viruses could provide virologists with an accurate means of viral identification and characterization. The aim of our work is to develop new methods for understanding local and global viral structure.