tetano
Editor, Senior Moderator
Antiviral Res
. 2020 Jul 9;104873.
doi: 10.1016/j.antiviral.2020.104873. Online ahead of print.
Characterization of Heparin and Severe Acute Respiratory Syndrome-Related Coronavirus 2 (SARS-CoV-2) Spike Glycoprotein Binding Interactions
So Young Kim[SUP] 1 [/SUP], Weihua Jin[SUP] 2 [/SUP], Amika Sood[SUP] 3 [/SUP], David W Montgomery[SUP] 4 [/SUP], Oliver C Grant[SUP] 4 [/SUP], Mark M Fuster[SUP] 5 [/SUP], Li Fu[SUP] 2 [/SUP], Jonathan S Dordick[SUP] 6 [/SUP], Robert J Woods[SUP] 4 [/SUP], Fuming Zhang[SUP] 7 [/SUP], Robert J Linhardt[SUP] 8 [/SUP]
Affiliations
Abstract
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 spike bind more tightly to immobilized heparin (K[SUB]D[/SUB] = 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 IC[SUB]50[/SUB] 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 μM, 0.12 μM, and 26.4 μM, 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.
Keywords: COVID-19; SARS-CoV-2; binding interactions; glycosaminoglycans; heparin; spike glycoprotein.
. 2020 Jul 9;104873.
doi: 10.1016/j.antiviral.2020.104873. Online ahead of print.
Characterization of Heparin and Severe Acute Respiratory Syndrome-Related Coronavirus 2 (SARS-CoV-2) Spike Glycoprotein Binding Interactions
So Young Kim[SUP] 1 [/SUP], Weihua Jin[SUP] 2 [/SUP], Amika Sood[SUP] 3 [/SUP], David W Montgomery[SUP] 4 [/SUP], Oliver C Grant[SUP] 4 [/SUP], Mark M Fuster[SUP] 5 [/SUP], Li Fu[SUP] 2 [/SUP], Jonathan S Dordick[SUP] 6 [/SUP], Robert J Woods[SUP] 4 [/SUP], Fuming Zhang[SUP] 7 [/SUP], Robert J Linhardt[SUP] 8 [/SUP]
Affiliations
- PMID: 32653452
- DOI: 10.1016/j.antiviral.2020.104873
Abstract
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 spike bind more tightly to immobilized heparin (K[SUB]D[/SUB] = 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 IC[SUB]50[/SUB] 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 μM, 0.12 μM, and 26.4 μM, 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.
Keywords: COVID-19; SARS-CoV-2; binding interactions; glycosaminoglycans; heparin; spike glycoprotein.