Characterization of heparin and severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) spike glycoprotein binding interactions
Characterization of heparin and severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) spike glycoprotein binding interactions
复制标题
DOI：
10.1016/j.antiviral.2020.104873
复制
发表时间：
2020-09-01
影响因子：
7.6
通讯作者：
Linhardt, Robert J.
中科院分区：
文献类型：
--
作者：
Kim, So Young;Jin, Weihua;Linhardt, Robert J.
严重急性呼吸综合征相关冠状病毒2（SARS-CoV-2）已导致大流行，并继续以前所未有的速度在地球仪各地传播。迄今为止，没有有效的治疗方法可用于对抗其相关疾病COVID-19。我们在SARS-CoV-2刺突糖蛋白（SGP）中发现了一个新的插入糖胺聚糖（GAG）结合基序（681-686（PRRARS））和另外两个GAG结合样基序，这使我们假设宿主细胞表面GAG可能与SARS-CoV-2 SGP相互作用以促进宿主细胞进入。使用表面等离子体共振直接结合试验，我们发现单体和三聚体SARS-CoV-2 SGP与固定化肝素的结合更紧密（KD 1/4分别为40 pM和73 pM），而SARS-CoV和MERS-CoV SGP（分别为500 nM和1 nM）。在竞争性结合研究中，肝素、三硫酸化非抗凝剂硫酸乙酰肝素和非抗凝剂低分子量肝素对SARS-CoV-2 SGP与固定化肝素结合的IC 50分别为0.056 μ M、0.12 μ M和26.4 μ M。最后，无偏计算配体对接表明硫酸乙酰肝素与GAG结合基序在三聚体SARS-CoV-2 SGP中每个单体界面上的S1/S2位点相互作用，并且当受体结合结构域处于开放构象时，硫酸乙酰肝素在另一个位点（453-459（YRLFRKS））相互作用。本研究为进一步研究糖胺聚糖在SARS-CoV-2发病机制中的生物学作用奠定了基础。此外，我们的研究结果可能为出现血栓性并发症的COVID-19患者的进一步基于肝素的干预提供额外的基础。
Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) has resulted in a pandemic and continues to spread around the globe at an unprecedented rate. To date, no effective therapeutic is available to fight its associated disease, COVID-19. Our discovery of a novel insertion of glycosaminoglycan (GAG)-binding motif at S1/S2 proteolytic cleavage site (681-686 (PRRARS)) and two other GAG-binding-like motifs within SARS-CoV-2 spike glycoprotein (SGP) led us to hypothesize that host cell surface GAGs may interact SARS-CoV-2 SGPs to facilitate host cell entry. Using a surface plasmon resonance direct binding assay, we found that both monomeric and trimeric SARS-CoV-2 SGP bind more tightly to immobilized heparin (KD 1/4 40 pM and 73 pM, respectively) than the SARS-CoV and MERS-CoV SGPs (500 nM and 1 nM, respectively). In competitive binding studies, the IC50 of heparin, tri-sulfated non-anticoagulant heparan sulfate, and non-anticoagulant low molecular weight heparin against SARS-CoV-2 SGP binding to immobilized heparin were 0.056 mu M, 0.12 mu M, and 26.4 mu M, respectively. Finally, unbiased computational ligand docking indicates that heparan sulfate interacts with the GAG-binding motif at the S1/S2 site on each monomer interface in the trimeric SARS-CoV-2 SGP, and at another site (453-459 (YRLFRKS)) when the receptor-binding domain is in an open conformation. The current study serves a foundation to further investigate biological roles of GAGs in SARS-CoV-2 pathogenesis. Furthermore, our findings may provide additional basis for further heparin-based interventions for COVID-19 patients exhibiting thrombotic complications.