{"doi":"10.1101/2020.05.21.109157","title":"Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity","abstract":"<jats:title>Abstract</jats:title>\n                <jats:p>A serious public health crisis is currently unfolding due to the SARS-CoV-2 pandemic. SARS-CoV-2 viral entry depends on an interaction between the receptor binding domain of the trimeric viral Spike protein (Spike-RBD) and the dimeric human angiotensin converting enzyme 2 (ACE2) receptor. While it is clear that strategies to block the Spike/ACE2 interaction are promising as anti-SARS-CoV-2 therapeutics, our current understanding is insufficient for the rational design of maximally effective therapeutic molecules. Here, we investigated the mechanism of Spike/ACE2 interaction by characterizing the binding affinity and kinetics of different multimeric forms of recombinant ACE2 and Spike-RBD domain. We also engineered ACE2 into a split Nanoluciferase-based reporter system to probe the conformational landscape of Spike-RBDs in the context of the Spike trimer. Interestingly, a dimeric form of ACE2, but not monomeric ACE2, binds with high affinity to Spike and blocks viral entry in pseudotyped virus and live SARS-CoV-2 virus neutralization assays. We show that dimeric ACE2 interacts with an RBD on Spike with limited intra-Spike avidity, which nonetheless contributes to the affinity of this interaction. Additionally, we demonstrate that a proportion of Spike can simultaneously interact with multiple ACE2 dimers, indicating that more than one RBD domain in a Spike trimer can adopt an ACE2-accessible “up” conformation. Our findings have significant implications on the design strategies of therapeutic molecules that block the Spike/ACE2 interaction. The constructs we describe are freely available to the research community as molecular tools to further our understanding of SARS-CoV-2 biology.</jats:p>","journal":null,"year":null,"id":638784,"datarank":0.5897738449086489,"base_score":3.9318256327243257,"endowment":3.9318256327243257,"self_citation_contribution":0.5897738449086489,"citation_network_contribution":0.0,"self_endowment_contribution":0.5897738449086489,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":50,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":237518,"name":"Xin Zhou","orcid":"0000-0002-8198-2856","position":1,"is_corresponding":false},{"id":237517,"name":"Shion A. Lim","orcid":"0000-0003-2136-2732","position":2,"is_corresponding":false},{"id":303659,"name":"Susanna K. Elledge","orcid":"0000-0002-9621-3881","position":3,"is_corresponding":false},{"id":239162,"name":"Paige Solomon","orcid":null,"position":4,"is_corresponding":false},{"id":237513,"name":"Nicholas J. Rettko","orcid":"0000-0002-4332-2697","position":5,"is_corresponding":false},{"id":227696,"name":"Beth Shoshana Zha","orcid":"0000-0002-4190-2823","position":6,"is_corresponding":false},{"id":292905,"name":"Lisa L. Kirkemo","orcid":"0000-0003-1686-6987","position":7,"is_corresponding":false},{"id":841672,"name":"Josef A. Gramespacher","orcid":"0000-0003-3478-0183","position":8,"is_corresponding":false},{"id":334282,"name":"Jia Liu","orcid":"0000-0001-9522-6624","position":9,"is_corresponding":false},{"id":16973,"name":"Frauke Muecksch","orcid":"0000-0002-0132-5101","position":10,"is_corresponding":false},{"id":1659378,"name":"Julio Cesar Cetrulo Lorenzi","orcid":null,"position":11,"is_corresponding":false},{"id":104730,"name":"Fabian Schmidt","orcid":"0000-0001-7731-6685","position":12,"is_corresponding":false},{"id":16974,"name":"Yiska Weisblum","orcid":"0000-0002-9249-1745","position":13,"is_corresponding":false},{"id":104703,"name":"Davide F. Robbiani","orcid":"0000-0001-7379-3484","position":14,"is_corresponding":false},{"id":104187,"name":"Michel C. Nussenzweig","orcid":"0000-0003-0592-8564","position":15,"is_corresponding":false},{"id":619511,"name":"Theodora Hatziioannou","orcid":null,"position":16,"is_corresponding":false},{"id":16992,"name":"Paul D. Bieniasz","orcid":"0000-0002-2368-3719","position":17,"is_corresponding":false},{"id":1659382,"name":"Oren S. Rosenburg","orcid":null,"position":18,"is_corresponding":false},{"id":237516,"name":"Kevin Leung","orcid":"0000-0002-2087-4974","position":19,"is_corresponding":false},{"id":237520,"name":"James A. Wells","orcid":"0000-0001-8267-5519","position":20,"is_corresponding":false},{"id":237510,"name":"Irene Lui","orcid":"0000-0002-6171-5443","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity","abstract":"<jats:title>Abstract</jats:title>\n                <jats:p>A serious public health crisis is currently unfolding due to the SARS-CoV-2 pandemic. SARS-CoV-2 viral entry depends on an interaction between the receptor binding domain of the trimeric viral Spike protein (Spike-RBD) and the dimeric human angiotensin converting enzyme 2 (ACE2) receptor. While it is clear that strategies to block the Spike/ACE2 interaction are promising as anti-SARS-CoV-2 therapeutics, our current understanding is insufficient for the rational design of maximally effective therapeutic molecules. Here, we investigated the mechanism of Spike/ACE2 interaction by characterizing the binding affinity and kinetics of different multimeric forms of recombinant ACE2 and Spike-RBD domain. We also engineered ACE2 into a split Nanoluciferase-based reporter system to probe the conformational landscape of Spike-RBDs in the context of the Spike trimer. Interestingly, a dimeric form of ACE2, but not monomeric ACE2, binds with high affinity to Spike and blocks viral entry in pseudotyped virus and live SARS-CoV-2 virus neutralization assays. We show that dimeric ACE2 interacts with an RBD on Spike with limited intra-Spike avidity, which nonetheless contributes to the affinity of this interaction. Additionally, we demonstrate that a proportion of Spike can simultaneously interact with multiple ACE2 dimers, indicating that more than one RBD domain in a Spike trimer can adopt an ACE2-accessible “up” conformation. Our findings have significant implications on the design strategies of therapeutic molecules that block the Spike/ACE2 interaction. The constructs we describe are freely available to the research community as molecular tools to further our understanding of SARS-CoV-2 biology.</jats:p>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"19910364","pmcid":null,"openalex_id":null,"authors":[],"funders":[],"total_grants":0,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"green","license":"cc-by-nc-nd","oa_locations":[{"url":"https://www.biorxiv.org/content/biorxiv/early/2020/05/21/2020.05.21.109157.full.pdf","host_type":"repository"},{"url":"https://syndication.highwire.org/content/doi/10.1101/2020.05.21.109157","host_type":"publisher"},{"url":"https://europepmc.org/article/PPR/PPR166043","host_type":"Europe_PMC"},{"url":"https://europepmc.org/api/fulltextRepo?pprId=PPR166043&type=FILE&fileName=EMS93632-pdf.pdf&mimeType=application/pdf","host_type":"Europe_PMC"}],"fields_of_study":[],"mesh_terms":[],"keywords":[],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"pdb"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-06T21:30:31.375807Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}