Biophysical studies of viral membrane fusion proteins

病毒膜融合蛋白的生物物理学研究

• 批准号: 10798382
• 负责人: James B Munro
• 金额: $ 5.78万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798382
• 关键词: 2019-nCoV; Address; Administrative Supplement; Biological Assay; Biological Models; Cell surface; Cells; Cellular Membrane; Chimeric Proteins; Cryo-electron tomography; Custom; Data; Endocytosis; Ensure; Environment; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Funding; Glycoproteins; Hemagglutinin; Image; Influenza; Influenza A virus; Influenza Hemagglutinin; Institution; Laboratories; Life Cycle Stages; Mediating; Membrane Fusion; Microscope; Molecular Conformation; Research; Sialic Acids; Surface; Time; Viral; Virion; Virus; biophysical analysis; design; endosome membrane; equipment acquisition; experimental study; late endosome; medical schools; premature; single molecule

SUMMARY Hemagglutinin (HA) is an envelope glycoprotein that resides on the surface of influenza A virus (IAV) particles. HA mediates attachment of IAV virions to the cell surface through interaction with sialic acid (SA) moieties. Following internalization by endocytosis, the acidic environment of the late endosome triggers conversion of HA from a metastable prefusion conformation to a highly stable postfusion coiled coil. This cascade of conformational changes in HA results in fusion of the viral and endosomal membranes. Importantly, exposure of IAV to acidic pH prior to attachment to a cellular membrane prematurely converts HA to the postfusion conformation, which renders IAV noninfectious. Therefore, ensuring proper timing of HA conformational changes is critical to the life cycle of IAV. While endpoints of the HA conformational changes during membrane fusion are well characterized, recent structural data is beginning to identify intermediate states. However, the HA conformational trajectory that leads from prefusion to postfusion is not yet validated, nor has the timing of membrane fusion with respect to HA conformation been determined. The funded project leverages the combined strengths of the Munro and Somasundaran laboratories in cutting-edge single-molecule Förster resonance energy transfer (smFRET) imaging, single-virion fusion assays, cryoelectron tomography (cryoET), and computational approaches to address these questions in viral fusion. The funded project relies heavily on the application of advanced, quantitative fluorescence microscopy within the Munro laboratory. With institutional support from UMass Chan Medical School, the Munro laboratory has custom built a microscope specifically designed to conduct the proposed experiments. The requested Administrative Supplement for Equipment Purchases will provide upgrades to the existing microscope. These upgrades will address shortcomings of the microscope, which limit the ability to successfully complete the proposed Aims, and which were not foreseen at the time of application.

总结

项目成果

hACE2-Induced Allosteric Activation in SARS-CoV versus SARS-CoV-2 Spike Assemblies Revealed by Structural Dynamics.

• DOI: 10.1021/acsinfecdis.3c00010
• 发表时间: 2023-06-09
• 期刊: ACS INFECTIOUS DISEASES
• 影响因子: 5.3
• 作者: Chen, Chengbo;Zhu, Richard;Hodge, Edgar A.;Diaz-Salinas, Marco A.;Nguyen, Adam;Munro, James B.;Lee, Kelly K. K.
• 通讯作者: Lee, Kelly K. K.