Heparin Inhibits Cellular Invasion by SARS-CoV-2: Structural Dependence of the Interaction of the Spike S1 Receptor-Binding Domain with Heparin
Heparin Inhibits Cellular Invasion by SARS-CoV-2: Structural Dependence of the Interaction of the Spike S1 Receptor-Binding Domain with Heparin
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DOI：
10.1055/s-0040-1721319
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发表时间：
2020-12-01
影响因子：
6.7
通讯作者：
Skidmore, Mark A.
中科院分区：
文献类型：
--
作者：
Mycroft-West, Courtney J.;Su, Dunhao;Skidmore, Mark A.
动物的发育和动态平衡依赖于数百种细胞外调节蛋白与胞外和胞外糖胺聚糖硫酸乙酰肝素(HS)的相互作用，许多微生物利用HS作为一种黏附和侵袭的手段。肝素是一种广泛使用的抗凝血药，在结构上与HS相似，是一种常见的实验替代品。外源性肝素可预防一系列病毒的感染，包括S冠状病毒分离株HSR1。在这里，我们表明，肝素抑制严重急性呼吸综合征冠状病毒-2(SARS-CoV-2)对Vero细胞的入侵高达80%，剂量可通过预防实现，尤其是在雾化可提供的范围内。表面等离子体共振和圆二色谱表明，肝素和依诺肝素是一种临床抗凝的低分子肝素，它与SARS-CoV-2的S_1蛋白受体结合域(S_1 RBD)结合并诱导构象变化。使用肝素衍生物和大小定义片段的文库来探索这种相互作用的结构基础。与RBD的结合更依赖于2-O或6-O硫酸盐基团的存在，而不是N-硫酸盐化，六糖是诱导RBD二级结构变化所需的最小尺寸。抑制病毒感染很可能是由于S1 RBD上肝素/HS与血管紧张素转换酶2的结合位点重叠所致。这些结果表明，通过改变肝素及其衍生物作为抗SARS-CoV-2和其他冠状病毒科成员的抗病毒药物，可以快速开发一线治疗药物。
The dependence of development and homeostasis in animals on the interaction of hundreds of extracellular regulatory proteins with the peri- and extracellular glycosaminoglycan heparan sulfate (HS) is exploited by many microbial pathogens as a means of adherence and invasion. Heparin, a widely used anticoagulant drug, is structurally similar to HS and is a common experimental proxy. Exogenous heparin prevents infection by a range of viruses, including S-associated coronavirus isolate HSR1. Here, we show that heparin inhibits severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) invasion of Vero cells by up to 80% at doses achievable through prophylaxis and, particularly relevant, within the range deliverable by nebulisation. Surface plasmon resonance and circular dichroism spectroscopy demonstrate that heparin and enoxaparin, a low-molecular-weight heparin which is a clinical anticoagulant, bind and induce a conformational change in the spike (S1) protein receptor-binding domain (S1 RBD) of SARS-CoV-2. A library of heparin derivatives and size-defined fragments were used to probe the structural basis of this interaction. Binding to the RBD is more strongly dependent on the presence of 2- O or 6- O sulfate groups than on N -sulfation and a hexasaccharide is the minimum size required for secondary structural changes to be induced in the RBD. It is likely that inhibition of viral infection arises from an overlap between the binding sites of heparin/HS on S1 RBD and that of the angiotensin-converting enzyme 2. The results suggest a route for the rapid development of a first-line therapeutic by repurposing heparin and its derivatives as antiviral agents against SARS-CoV-2 and other members of the Coronaviridae .