Molecular architecture of the Vaccinia virion by structural proteomics

通过结构蛋白质组学研究牛痘病毒粒子的分子结构

基本信息

批准号：
10179428
负责人：
Paul D Gershon
金额：
$ 33.2万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-22 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179428
关键词：
3-Dimensional ART protein Africa Agriculture Animals Antiviral Agents Appearance Architecture Biological Assay Capsid Capsid Proteins Cells Chemicals Complex Crosslinker DNA Polymerase II DNA Viruses DNA-Directed RNA Polymerase Data Data Set Disease Disease Outbreaks Docking Drug Targeting Ebola Enzymes Equipment and supply inventories Escherichia coli Exploratory/Developmental Grant for Diagnostic Cancer Imaging Family Family member Future Genes Genome Goals Grant Health Heart Heterogeneity Human Image Immune system In Situ Infection Infusion procedures Intervention Ions Isotope Labeling Knowledge Laboratories Light Linear Programming Mass Spectrum Analysis Mediating Medical Methods Molecular Molecular Structure Monkeypox Morphogenesis Nucleocapsid Pathway interactions Peptides Phenotype Poxviridae Preparation Process Proteins Proteomics Protomer Publishing RNA RNA Viruses Resolution Roentgen Rays Route Small RNA Smallpox Smallpox Vaccine Structural Models Structural Protein Structure Tertiary Protein Structure Testing Therapeutic Time Vaccination Vaccines Vaccinia Vaccinia virus Viral Virion Virus Virus Replication Yeasts ZIKA anthrax lethal factor base bioinformatics tool connectome crosslink design env Gene Products flexibility innovation interest molecular modeling monomer mutant particle protein crosslink protein folding protein structure self organization stoichiometry therapeutic development three dimensional structure tool transmission process virus envelope

项目摘要

The poxviruses comprise a major virus family of medical, ecological and agricultural importance. The most notorious family member, smallpox has been one of the great killers of mankind. Although the disease was eradicated some 40 years ago, the possibility of smallpox re- appearance at some future time has increased immeasurably with the recent demonstration that a poxvirus very similar to smallpox could be recreated de novo, in the laboratory, with ease. Moreover, eradication and the cessation of vaccination has coincided with the appearance of feral human poxviruses including human monkeypox in Africa, the US and UK. Not knowing the lethal factor in smallpox, the full potential of such outbreaks remains uncertain. The importance of virus envelope and capsid proteins in mediating the effects of antiviral therapeutics and vaccines is undisputed. For small RNA viruses in particular, an understanding of virion structure at molecular or atomic resolution has instructed the development of therapeutic agents and an understanding of mechanisms of infection and disease. Due to their complexity, asymmetry and heterogeneity, poxvirus virions have, however, persistently eluded attempts to elucidate their molecular structure, closing a potential avenue of rational design and intervention. The P.I. hypothesizes that the relative complexity of the vaccinia virion may be a therapeutic Achilles heel. Moreover, a molecular-level understanding of virion morphogenesis and organization, one of the last remaining black boxes in the lifecycle of the poxviruses, impinges upon at least five of the seven classical stages of virus replication. A major gap in our knowledge of pox virion structure lies at the level of molecular architecture – an intervening organizational level between ultrastructural features and the inventory of protein molecules contained within the virion. The P.I. has successfully applied a protein-protein chemical crosslinking approach in combination with protein mass spectrometry (XLMS) to discover neighboring proteins and domains within the undisrupted vaccinia virion in situ. Aim 1 of this proposal seeks to deepen the XLMS dataset to a level that will allow protein molecular docking. Combining XLMS with mutant virus particles blocked in morphogenesis and displaying no apparent internal organization, Aim 2 of this proposal asks whether the virion morphogenic pathway follows a classical programmed linear hierarchy or a process of self-organization with no single, dominant route from molecular components to assembled virion. Using “QconCAT” quantitative MS, Aim 3 seeks to convert XLMS data to a molecular model by determining the global stoichiometries of virion proteins.

痘病毒是具有医学、生态和农业重要性的主要病毒家族。天花是最臭名昭著的家族成员，一直是人类的大杀手之一。尽管这种疾病在大约 40 年前就已被根除，但天花的可能性仍然存在。随着最近的证明，未来某个时间的出现次数已大大增加与天花非常相似的痘病毒可以在实验室中轻松地从头重建。此外，根除和停止疫苗接种与新冠病毒的出现同时发生。野生人类痘病毒，包括非洲、美国和英国的人类猴痘病毒。天花的致命因素，这种爆发的全部潜力仍然不确定。病毒包膜和衣壳蛋白在介导抗病毒治疗效果中的作用尤其是对于小 RNA 病毒来说，对病毒颗粒结构的了解是毫无争议的。分子或原子分辨率指导了治疗剂和药物的开发了解感染和疾病的机制，因为它们的复杂性、不对称性和然而，痘病毒病毒粒子一直未能阐明其异质性分子结构，关闭了理性设计和干预的潜在途径。陷阱认为痘苗病毒颗粒的相对复杂性可能是治疗的致命弱点此外，对病毒粒子形态发生和组织的分子水平理解，之一。痘病毒生命周期中最后剩下的黑匣子，撞击了至少五个病毒复制的七个经典阶段是我们对痘病毒体知识的一个主要空白。结构位于分子结构的水平上——介于两者之间的介入组织水平超微结构特征和病毒颗粒内所含蛋白质分子的库存。 P.I. 成功应用了蛋白质-蛋白质化学交联方法使用蛋白质质谱 (XLMS) 来发现邻近的蛋白质和结构域该提案的目标 1 旨在加深 XLMS 数据集以实现原位未破坏的痘苗病毒粒子。允许蛋白质分子对接的水平。形态发生受阻，没有明显的内部组织，目标 2 该提案询问病毒粒子形态发生途径是否遵循经典的线性编程层次结构或自组织过程，没有来自分子的单一主导途径 Aim 3 试图使用“QconCAT”定量 MS 将组件转化为组装的病毒颗粒。通过确定病毒体蛋白的整体化学计量，将 XLMS 数据转化为分子模型。