Effect of natural and engineered variations on structure and biophysics of SARS-CoV-2 spike

自然和工程变异对 SARS-CoV-2 刺突结构和生物物理学的影响

基本信息

批准号：
10453964
负责人：
Priyamvada Acharya
金额：
$ 76.25万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10453964
关键词：
2019-nCoV Antibodies Antigens Biochemical Biochemistry Biological Biophysics COVID-19 Communication Computing Methodologies Coronavirus Coronavirus spike protein Cryoelectron Microscopy Data Data Set Electron Microscopy Emergency Situation Engineering Evolution Fc Receptor Funding Future Goals Grant Immune Evasion In Vitro Kinetics Lead Life Cycle Stages Link Measures Methods Mink Molecular Molecular Conformation Mutagenesis Mutation Negative Staining Pathogenesis Peptide Hydrolases Peptides Population Positioning Attribute Predisposition Property Protein Conformation Proteins Proteolysis Protomer Recurrence Reporting Research Resistance Resolution Role SARS-CoV-2 B.1.1.7 SARS-CoV-2 B.1.351 SARS-CoV-2 B.1.617.2 Site Structure Surface TMPRSS2 gene Techniques Technology United States National Institutes of Health Vaccine Design Vaccines Variant Viral Virus Work X-Ray Crystallography advanced simulation base betacoronavirus biophysical analysis combat computer studies design expectation experimental study global health health economics in vivo innovation insight neutralizing antibody novel vaccines pandemic disease protein structure receptor binding reconstruction structural biology vaccine development vaccine hesitancy variants of concern

项目摘要

Effect of natural and engineered variations on structure and biophysics of SARS-CoV-2 spike COVID-19, caused by SARS-CoV-2, has devasted global health and economics. Vaccines are being deployed worldwide to gain control of the pandemic, although emergence of fast-spreading “variants of concern” (VOCs) have caused concern. Mutations in the spike (S) protein are under scrutiny due to its essential role in the virus life cycle, and being the dominant target of neutralizing antibodies. Widespread vaccine hesitancy and the current spread of the Delta variant provide fertile ground for emergence of vaccine- resistant variants. We and others have shown that variants use a plethora of strategies to modify antibody and receptor interactive surfaces, and spike conformation, resulting in antibody evasion and greater infectivity. Over the last two years, utilizing urgent supplement funding from the NIH, we studied the structures of SARS- CoV-2 S proteins and have established workflows spanning structure, biochemistry, biophysics and computation. Here we propose to continue the essential work of detangling the effects of variant S protein mutations, and to enhance our understanding of spike structure to further efforts to predict where the virus is heading and to inform novel vaccine designs. The scientific premise of this grant is that understanding spike structure and allostery will provide insights into its function, inform vaccine development, and provide mechanistic information essential for relating spike structure to beta-CoV replication, evolution, and immune evasion. The innovations in this grant derive from technologies we have developed for structural analyses of the S protein: an integrative structural biology pipeline combines cryo-electron microscopy (cryo-EM), Negative Stain Electron Microscopy (NSEM) and X-ray crystallography, with computational methods, and biochemical and biophysical analyses to study structural and functional properties of the spike, including furin cleavage, receptor binding, and antigenicity.

自然和工程变化对SARS-COV-2 SPIKE的结构和生物物理学的影响由SARS-COV-2引起的Covid-19毁了全球健康和经济学。正在疫苗尽管出现快速发展的“变种” 关注”（VOC）引起了人们的关注。峰值蛋白的突变由于其蛋白质而受到审查在病毒生命周期中的重要作用，并且是中和抗体的主要靶标。广泛疫苗的犹豫和三角洲变体的当前扩散为疫苗出现提供肥沃的地面 - 抗性变体。我们和其他人表明，变体使用多种策略来修改抗体和受体相互作用表面和尖峰构象，导致抗体演化和更大的感染。在过去的两年中，使用NIH的紧急补充资金，我们研究了SARS- COV-2 S蛋白质并建立了跨越结构，生物化学，生物物理学和计算。在这里，我们建议继续缠绕变体S蛋白的影响的基本工作突变，并增强我们对峰值结构的理解，以进一步预测病毒在哪里标题并为新颖的疫苗设计提供信息。这笔赠款的科学前提是理解Spike 结构和变构将提供有关其功能的见解，为疫苗开发提供信息，并提供将尖峰结构与β-COV复制，进化和免疫相关的机械信息这笔赠款的创新源自我们为结构分析而开发的技术 S蛋白：综合结构生物学管道结合了冷冻电子显微镜（Cryo-EM），阴性染色电子显微镜（NSEM）和X射线晶体学，具有计算方法，生化方法和生物物理分析以研究尖峰的结构和功能特性，包括脂蛋白裂解，受体结合和抗原性。