Multimode Observation of Virus Capsid Assembly

病毒衣壳组装的多模式观察

基本信息

批准号：
10587218
负责人：
Adam Zlotnick
金额：
$ 49.34万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-14 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10587218
关键词：
Address Affinity Allosteric Regulation Antibodies Antiviral Agents Behavior Binding Biochemistry Biological Assay Biophysics Biotechnology Capsid Capsid Proteins Cells Charge Chemicals Chemistry Chronic Chronic Hepatitis B Complex Crowding Cytoplasm DNA biosynthesis Data Detection Development Devices Dissociation Endosomes Engineering Environment Epitopes Exposure to Funding Genome Goals Growth Hepatitis B Virus Kinetics Learning Life Cycle Stages Ligand Binding Link Mass Spectrum Analysis Measures Metabolic Methods Modeling Modification Molecular Biology Oxidation-Reduction Paper Penetration Peptides Proteins RNA Reaction Reagent Research Roentgen Rays Role Self Correction Signal Transduction Structure Surface System Techniques Technology Testing Text Thermodynamics Time Variant Viral Viral Genome Viral Packaging Virus Virus Assembly Virus Diseases Virus-like particle delivery vehicle dimer falls flexibility human pathogen improved in vivo macromolecule man particle exposure practical application preservation protein purification receptor receptor binding response seal self assembly single molecule small molecule tool viral RNA

项目摘要

Summary The capsid of the Hepatitis B Virus (HBV) is a 120-homodimer T=4 icosahedron. In vivo, it self-assembles, packages viral RNA, serves as a metabolic compartment for DNA synthesis, and trafficks within the cell. Its assembly and disassembly have become targets for potent antivirals. In the previous funding period, we characterized assembly and disassembly with purified protein using structural, single molecule, and bulk studies of assembly products and reactions. We characterized the allosteric transitions that activated assembly, demonstrated the importance of reversibility during self-association for fidelity, and identified roles of nucleation for directing the assembly path. In this proposal, we develop hypotheses to take advantage of these results to engineer virus-like particles with programmable assembly, cargo packaging, delivery, and release. In preliminary studies, we developed a method for targeting cargo to a capsid by linking the cargo to a small molecule that binds capsid with high affinity, essentially using an antiviral as a targeting device. This method can be applied to any cargo. In some cases, it is desirable to display cargo on the capsid exterior, in other cases it is desirable to package it within the capsid. In preliminary data, we demonstrate an approach to making “holey” capsids that expose the particle interior and can be re-sealed to enclose the contents. Using this same technology, we can make patches on the capsid surface; this can be used for displaying patches of receptor-binding ligands or cell-penetrating peptides. Cargo, packaged within a capsid, is not deliverable unless it can be released. In preliminary data, we developed techniques for triggering capsid disassembly in response to redox potential, taking advantage of chemically-induced metastability. This same approach can be applied to other triggering signals. The ultimate goal of these studies is to combine the approaches to a practical end: we propose to build two model biotech reagents, one to measure antibody levels and the other to deliver packaged cargo to specific cells. These approaches are each built on an understanding of the biochemistry and biophysics of HBV capsid assembly. HBV is one the smallest human pathogens. It is remarkably efficient at packaging its genome and delivering it to target cells. Based on our understanding of capsid assembly and capsid biophysics, we will develop approaches to specifically packaging cargo molecules and delivering these reagents intracellularly. The tools arising from this research will provide a means for man to take advantage of HBV.

概括丙型肝炎病毒（HBV）的衣壳是120-二二聚体T = 4二十面体。在体内，它可以自组成，包装病毒RNA，是用于DNA合成的代谢室和细胞内的交通。它的组装和拆卸已成为潜在抗病毒药的靶标。在上一个资金期间，我们使用结构，单分子和组装产品和反应的批量研究。我们表征了激活的变构转变组装，证明了在自我关联过程中可逆性的重要性，并确定成核指导组装路径的作用。在此提案中，我们提出假设以利用这些结果来设计类似病毒的颗粒具有可编程组件，货物包装，交付和释放。在初步研究中，我们通过将货物与结合的小分子联系起具有高亲和力的衣壳，基本上使用抗病毒作为靶向装置。此方法可以应用于任何货物。在某些情况下，希望在衣壳外部显示货物，在其他情况下是可取的将其包装在Capsid中。在初步数据中，我们演示了一种制作“洞”帽质的方法暴露于粒子内部并可以重新密封以包围内容。使用相同的技术，我们可以在衣壳表面上制作斑块。这可用于显示接收器结合的补丁配体或细胞穿透肽。货物，包装在衣壳中，除非可以发行。在初步数据中，我们开发了用于触发capsid拆卸的技术氧化还原电位，利用化学诱导的亚稳定性。可以采用相同的方法到其他触发信号。这些研究的最终目标是结合实用的方法结束：我们建议建立两种模型生物技术试剂，一种用于测量抗体水平，另一个用于将包装的货物运送到特定的细胞。这些方法都建立在对 HBV衣壳组件的生物化学和生物物理学。 HBV是最小的人类病原体之一。它在包装基因组和交付方面非常有效它是靶细胞。根据我们对衣壳组装和衣壳生物物理学的理解，我们将发展专门包装货物分子并细胞内传递这些试剂的方法。这这项研究产生的工具将为人类利用HBV提供一种手段。

项目成果

期刊论文数量（25）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hepatitis B virus core protein allosteric modulators can distort and disrupt intact capsids.

DOI：
10.7554/elife.31473
发表时间：
2018-01-29
期刊：
eLife
影响因子：
7.7
作者：
Schlicksup CJ;Wang JC;Francis S;Venkatakrishnan B;Turner WW;VanNieuwenhze M;Zlotnick A
通讯作者：
Zlotnick A

The Structural Biology of Hepatitis B Virus: Form and Function.