Structural Studies of Coronavirus Fusion Proteins

冠状病毒融合蛋白的结构研究

基本信息

批准号：
9324294
负责人：
David Veesler
金额：
$ 29万
依托单位：
UNIVERSITY OF WASHINGTON
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：
2016-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9324294
关键词：
Address Antibodies Antigens Architecture Attention Binding Biological Assay Biological Models Cell fusion Cell membrane Cell-Matrix Junction Cells Chimeric Proteins Complement Complex Coronaviridae Coronavirus Coronavirus Infections Coronavirus spike protein Cryoelectron Microscopy Data Data Analyses Disease Outbreaks Family Fostering Future Genetic Materials Genome Glycoproteins Goals Health Human Immunization Immunoglobulin Fragments Infection Kinetics Lead Mediating Membrane Membrane Fusion Middle East Respiratory Syndrome Coronavirus Modeling Molecular Molecular Conformation Murine hepatitis virus Mus Mutagenesis N-terminal Negative Staining Outcome Pathogenicity Pneumonia Process Protein Engineering Proteins Reaction Resolution Structure Surface Thermodynamics Viral Viral Fusion Proteins Virus Virus Diseases Work antibody engineering antibody inhibitor biophysical techniques coronavirus spike glycoprotein design experimental study grasp mutant neutralizing antibody pandemic disease particle prevent receptor receptor binding reconstruction small molecule inhibitor stem targeted treatment therapeutic target three-dimensional modeling vaccinology virus envelope

项目摘要

Enveloped viruses use specialized proteins present at the virus surface to translocate their genetic material across the host cell membrane during infection. For coronaviruses, homotrimers of the spike glycoprotein promote host cell attachment and fusion of the viral and host membranes. Although coronaviruses have a significant pandemic potential, the lack of high-resolution data for any coronavirus spike trimer limits our mechanistic understanding of infection by this family of viruses. The objective of the proposed work is to obtain high-resolution snapshots corresponding to the various stages of the fusion reaction mediated by coronavirus spikes and to study the structural determinants associated with antibody inhibition of viral infection. In Aim I, we propose to elucidate the architecture of the Mouse Hepatitis Virus (MHV) pre-fusion spike using cryoEM. Aim II will be dedicated to studying the conformational changes associated with the fusion reaction with an emphasis on the first intermediate (extended intermediate) and the post-fusion spike. In Aim III, we will characterize the 3D organization of human coronavirus spikes to understand how these viruses overcome the species barrier and to identify structurally conserved regions that could be potential targets for therapeutic initiatives. The final aim will rely on structure-guided protein design to engineer antibodies targeting human coronavirus spikes with the goal of identifying immunogens for raising broadly-neutralizing antibodies.

包裹的病毒使用存在于病毒表面的专门蛋白质来转移其遗传感染过程中跨宿主细胞膜的材料。对于冠状病毒，尖峰的同构二聚体糖蛋白促进病毒和宿主膜的宿主细胞附着和融合。虽然冠状病毒具有巨大的大流行潜力，对于任何冠状病毒尖峰三聚体限制了我们对这种病毒家族感染的机械理解。拟议工作的目的是获得对应于该快照的高分辨率快照冠状病毒尖峰介导的融合反应的各个阶段，并研究结构与抗体抑制病毒感染相关的决定因素。在目标一世中，我们建议阐明小鼠肝炎病毒（MHV）使用冷冻峰的结构。 AIM II将是致力于研究与融合反应相关的构象变化强调第一个中间体（扩展中间体）和融合后尖峰。在AIM III中，我们将表征人类冠状病毒尖峰的3D组织，以了解这些病毒克服物种障碍并确定可能是潜在的结构保守区域治疗计划的目标。最终目标将依靠结构引导的蛋白质设计来工程靶向人冠状病毒尖峰的抗体，目的是识别升高的免疫原分广泛中和抗体。