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.

冠状病毒是一种膜包膜，表面有突出的“刺突”，这些刺突与宿主细胞相互作用，引发该细胞的感染。冠状病毒传播到其他细胞的能力取决于刺突与其相互作用的程度。控制这种相互作用的刺突部分称为“融合肽”，其作用是插入宿主膜以启动将其基因货物递送到细胞中。与膜相互作用在这个项目中，将研究融合肽与模拟不同种类宿主细胞的膜表面之间的各种相互作用。这些信息将提供对融合肽和促进它们相互作用的宿主细胞的特征的深入了解。 ，以及最终，感染和病毒传播到新宿主。随着对病毒适应新宿主的这种关键相互作用背后的科学有了更好的了解，我们将更好地了解这种病毒是如何传播的，并最终有能力制定阻止它的策略。这一基本信息有可能为解决这一问题提供新的方法。设计针对融合肽的抗病毒药物。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COVID-19的冠状病毒。冠状病毒能力的关键决定因素对于冠状病毒来说，进入宿主细胞是由从其膜包膜突出的单个糖蛋白（称为刺突（S））介导的，在 S 内，直接与膜相互作用的区域是。被称为融合肽，FP。正是 FP 与宿主膜的物理化学相互作用将其固定，从而使膜发生必要的变形，从而将病毒基因组递送到细胞中。最基本的层面将有助于制定限制这些相互作用以阻止感染传播的策略，这一信息预计将有助于预测未来可能对人类构成威胁的新菌株的特征。项目的目的是测量和识别该项目将：1）利用原子力显微镜和生物膜模型的单分子力测量来阐明控制 FP 疏水相互作用的因素，以及 2）表征结构 -使用圆二色性和等温量热法将 FP 的功能关系与氨基酸序列和宿主表面特性关联起来，该项目的智力价值是发现宿主膜化学或氨基酸的变化。 FP 的序列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，该项目的更广泛影响是提供信息，使新的抗病毒药物设计方法成为可能，并确定可提供信息的基本设计规则。 FP 如何促进与特定化学膜的相互作用，以预测宿主对感染的易感性。鉴于 FP 在 CoV 家族中高度保守，这些研究将直接适用于所有 CoV，包括那些尚未出现的 CoV。该奖项反映了这一点。通过使用基金会的智力价值和更广泛的影响审查标准进行评估，NSF 的法定使命被认为值得支持。