RAPID: Revealing the intermolecular interactions between the SARS-CoV-2/COVID-19 fusion peptide and the host cell membrane that underlie its flexibility in host tropism

RAPID：揭示 SARS-CoV-2/COVID-19 融合肽与宿主细胞膜之间的分子间相互作用，这是其宿主向性灵活性的基础

• 批准号: 2027070
• 负责人: Susan Daniel
• 金额: $ 20万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2027070&HistoricalAwards=false
• 关键词: RAPID; Revealing; intermolecular; interactions; between

Coronavirus is a membrane-enveloped package decorated with “spikes” that protrude from its surface. These spikes interact with the host cell to initiate infection of that cell. The ability of coronavirus to spread to other cells depends on how well the spike interacts with its target host membrane. The part of the spike that controls this interaction is called the “fusion peptide” and its job is to insert into the host membrane to initiate the delivery its genetic cargo into the cell. However, the specifics of how the fusion peptide interacts with the membrane is still not known. In this project, various interactions between the fusion peptide with membrane surfaces that mimic different kinds of host cells will be studied. This information will provide insight into the characteristics of both the fusion peptide and the host cell that promotes their interaction, and ultimately, infection and viral transmission to new hosts. With a better understanding of the science behind this critical interaction for virus adaptation to new hosts, we will be better informed about how this virus spreads and ultimately equipped to develop strategies to stop it. This fundamental information has the potential to enable fresh approaches to the design of antiviral drugs that target the fusion peptide. Given that the fusion peptide is highly conserved across the coronavirus family, these studies will be directly applicable to all coronaviruses, including the coronavirus that causes COVID-19.A key determinant of the ability of coronavirus to spread is how it interacts with its target host membrane. For coronavirus, entry into a host cell is mediated by a single glycoprotein protruding from its membrane envelope, called spike (S). Within S, the region that directly interacts with the membrane is called the fusion peptide, FP. It is the physico-chemical interactions of the FP with the host membrane that anchors it, thus enabling the necessary deformations of the membrane that lead to delivery of the viral genome into the cell. Thus, understanding FP interactions at the most fundamental level will facilitate the development of strategies to limit those interactions to stop the spread of infections. This information is expected to be helpful in predicting the characteristics of emerging strains that could post a threat to humans in the future. The objective of this project is to measure and identify the specific intermolecular interactions responsible for insertion of FP into membranes. Specifically, this project will: 1) elucidate the factors that control hydrophobic interactions of FP using single molecule force measurements with atomic force microscopy and models of biological membranes, and 2) characterize the structure-function relationship of the FP using circular dichroism and isothermal calorimetry to correlate specific interactions with amino acid sequence and host surface properties. The intellectual merit of this project is discovering how changes in host membrane chemistry or amino acid sequence of the FP modulates the hydrophobic interaction and ultimately influences activity critical to the spread of infection. The broader impact of this project is providing information that will enable fresh approaches to the design of antiviral drugs, as well as to identify basic design rules that inform how the FP promotes the interaction with membranes of specific chemistry to predict host susceptibility to infection. Given that the FP is highly conserved across the CoV family, these studies will be directly applicable to all CoVs, including those yet to emerge.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

冠状病毒是一种由膜开发的包装，上面装饰有“尖峰”，从其表面伸出。这些尖峰与宿主细胞相互作用以引发该细胞的感染。冠状病毒扩散到其他细胞的能力取决于峰值与靶宿主膜的相互作用。控制这种相互作用的尖峰部分称为“融合肽”，其作业是将其插入宿主膜中以将其遗传货物传递到细胞中。但是，融合肽如何与膜相互作用的细节尚不清楚。在这个项目中，融合肽与膜表面之间的各种相互作用，模仿不同种类的宿主细胞将被研究。该信息将洞悉融合肽和促进其相互作用的宿主细胞的特征，并最终向新宿主传播感染和病毒传播。有了更好地了解这种对新宿主病毒适应新宿主的关键互动背后的科学，我们将更好地了解这种病毒如何传播，并最终等同于制定阻止它的策略。这些基本信息有可能使针对融合肽的抗病毒药物的设计新鲜方法。鉴于融合肽在冠状病毒家族中高度保守，这些研究将直接适用于所有冠状病毒，包括引起冠状病毒的冠状病毒，这是冠状病毒传播能力的关键确定剂，是它如何与靶宿主膜相互作用。对于冠状病毒，进入宿主细胞是由一种称为Spike（S）的单个糖蛋白蛋白介导的。在S中，直接与膜相互作用的区域称为融合肽FP。 FP与宿主膜的物理化学相互作用锚定，从而使膜的必要变形导致病毒基因组传递到细胞中。这样一来，了解最基本水平的FP相互作用将支持制定限制这些相互作用以阻止感染传播的策略。预计该信息将有助于预测可能在未来对人类威胁的新兴菌株的特征。该项目的目的是测量和确定负责将FP插入膜中的特定分子间相互作用。具体而言，该项目将：1）使用单分子力量测量与原子力显微镜和生物学机制模型来控制FP的疏水相互作用的因素，以及2）使用圆形二进制症和等渗性钙列列术与具有分离型宿主表面的特异性相互作用来表征FP的结构 - 功能关系。该项目的智力优点是发现FP的宿主膜化学或氨基酸序列的变化如何调节疏水相互作用，并最终影响对感染传播至关重要的活性。该项目的更广泛的影响是提供信息，可以使抗病毒药的设计新颖方法，以及确定基本的设计规则，以告知FP如何促进与特定化学机制的相互作用，从而预测宿主的感染易感性。鉴于FP在COV家族中是高度保守的，因此这些研究将直接适用于所有COV，包括尚未出现的COV。该奖项反映了NSF的法定任务，并通过使用该基金会的知识分子优点和更广泛的影响来审查标准，被视为通过评估来获得支持。