Dynamics and mechanisms of filovirus envelop glycoproteins

丝状病毒包膜糖蛋白的动力学和机制

基本信息

批准号：
10707317
负责人：
James B Munro
金额：
$ 78.28万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
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：
2022-09-19 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707317
关键词：
Adopted Antigens Binding Biophysics Cells Cellular Membrane Chemicals Cues Cysteine Proteinase Inhibitors Data Dependence Disease Outbreaks Ebola Ebola virus Electrostatics Endosomes Environment Event Excision Filovirus Fluorescence Frequencies GP2 gene Glycoproteins Goals Integration Host Factors Interdisciplinary Study Lipids Marburgvirus Mediating Membrane Membrane Fusion Methods Modeling Molecular Molecular Conformation Mucins Peptide Hydrolases Phenotype Phospholipids Physiological Play Polysaccharides Preventive measure Process Productivity Proteins Proteolysis Publications Publishing Research Role Severities Specific qualifier value Structure Surface Testing Therapeutic antibodies Vaccines Viral Virion Virus Virus Diseases Work biophysical techniques combat inhibitor insight late endosome multidisciplinary novel therapeutics receptor receptor binding response single molecule single-molecule FRET stem success

项目摘要

SUMMARY The increasing frequency and severity of Ebola outbreaks demands an expanded repertoire of treatments and preventative measures. Answering this unmet need will require a deeper understanding of the molecular mechanisms underlying filovirus replication. In particular, the dynamic events that occur during filovirus envelope glycoprotein (GP)-mediated membrane fusion during entry into cells have evaded elucidation for decades. Previous studies have identified proteolytic cleavage of GP, receptor binding, and the chemical environment of the late endosome as being critical. But the molecular mechanisms by which these events and variables promote GP-mediated membrane fusion are not known. As a result, a complete and specific model of filovirus fusion, which integrates host factors, environmental conditions, and GP conformational changes currently does not exist. Therefore, filovirus entry continues to be an unutilized target for inhibitors. Our long-term goal is to develop a complete mechanistic model of GP-mediated membrane fusion. Our recent publications, in which we demonstrate the power of single-molecule fluorescence methods in elucidating the conformational dynamics of EBOV GP on the surface of virions, demonstrate our initial efforts toward this end. Here we aim to build on this success by proposing a multidisciplinary study involving virological, cellular, structural, and biophysical methodologies to elucidate the dynamics and mechanisms of GPs from multiple filoviruses. We will characterize the mechanisms by which conformational changes, host factors, and environmental variables facilitate filovirus membrane fusion and entry into cells. Completion of the proposed research will provide mechanistic insights into filovirus entry and the viral and host targets that could be exploited with novel therapies and immunogens.

概括埃博拉疫情爆发的频率和严重程度的增加，需要扩大的治疗方法和预防措施。回答这种未满足的需求将需要更深入地了解分子丝状病毒复制的机制。特别是，在丝状病毒信封期间发生的动态事件进入细胞期间糖蛋白（GP）介导的膜融合已逃避了数十年。先前的研究已经确定了GP，受体结合和化学环境的蛋白水解切割晚期内体非常关键。但是这些事件和变量促进的分子机制 GP介导的膜融合尚不清楚。结果，一个完整而特定的filovirus融合模型，当前不存在整合宿主因素，环境条件和GP构象变化。因此，FILOVIRUS进入仍然是抑制剂的未利用靶标。我们的长期目标是开发 GP介导的膜融合的完整机械模型。我们最近的出版物，我们演示单分子荧光方法在阐明构象动力学方面的力量 Ebov GP在病毒体表面，展示了我们在这一目标上的最初努力。在这里我们旨在以此为基础通过提出一项涉及病毒学，细胞，结构和生物物理的多学科研究来取得成功阐明来自多个FILOVIRES的GPS的动力学和机制的方法。我们将表征构象变化，宿主因子和环境变量的机制有助于丝状病毒膜融合并进入细胞。拟议研究的完成将提供机械见解铁病毒的进入以及可以用新型疗法和免疫原分来利用的病毒和宿主靶标。