Structural Studies of Coronavirus Fusion Proteins

冠状病毒融合蛋白的结构研究

• 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
• 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

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.

有包膜病毒利用病毒表面的特殊蛋白质来转移它们的基因， 在感染期间穿过宿主细胞膜的物质。对于冠状病毒，刺突的同源三聚体 糖蛋白促进宿主细胞附着以及病毒和宿主膜的融合。虽然 冠状病毒具有重大的大流行潜力，缺乏任何高分辨率数据， 冠状病毒刺突三聚体限制了我们对该病毒家族感染的机制理解。 所提出的工作的目标是获得高分辨率快照对应的 冠状病毒刺突介导的融合反应的各个阶段，并研究其结构 与抗体抑制病毒感染相关的决定因素。在目的I中，我们建议澄清 使用cryoEM的小鼠肝炎病毒（MHV）融合前刺突的结构。目标二是 致力于研究与融合反应相关的构象变化， 强调第一中间体（延伸中间体）和融合后尖峰。在Aim III中，我们 将描述人类冠状病毒尖峰的3D组织，以了解这些病毒如何 克服物种障碍，并确定结构保守的区域， 治疗计划的目标。最终的目标将依赖于结构导向的蛋白质设计， 针对人类冠状病毒刺突的抗体，目的是鉴定用于提高 广泛中和抗体