{"doi":"10.1371/journal.ppat.1009519","title":"Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity","abstract":"SARS-CoV-2 is the novel coronavirus that is the causative agent of COVID-19, a sometimes-lethal respiratory infection responsible for a world-wide pandemic. The envelope (E) protein, one of four structural proteins encoded in the viral genome, is a 75-residue integral membrane protein whose transmembrane domain exhibits ion channel activity and whose cytoplasmic domain participates in protein-protein interactions. These activities contribute to several aspects of the viral replication-cycle, including virion assembly, budding, release, and pathogenesis. Here, we describe the structure and dynamics of full-length SARS-CoV-2 E protein in hexadecylphosphocholine micelles by NMR spectroscopy. We also characterized its interactions with four putative ion channel inhibitors. The chemical shift index and dipolar wave plots establish that E protein consists of a long transmembrane helix (residues 8-43) and a short cytoplasmic helix (residues 53-60) connected by a complex linker that exhibits some internal mobility. The conformations of the N-terminal transmembrane domain and the C-terminal cytoplasmic domain are unaffected by truncation from the intact protein. The chemical shift perturbations of E protein spectra induced by the addition of the inhibitors demonstrate that the N-terminal region (residues 6-18) is the principal binding site. The binding affinity of the inhibitors to E protein in micelles correlates with their antiviral potency in Vero E6 cells: HMA ≈ EIPA > DMA >> Amiloride, suggesting that bulky hydrophobic groups in the 5' position of the amiloride pyrazine ring play essential roles in binding to E protein and in antiviral activity. An N15A mutation increased the production of virus-like particles, induced significant chemical shift changes from residues in the inhibitor binding site, and abolished HMA binding, suggesting that Asn15 plays a key role in maintaining the protein conformation near the binding site. These studies provide the foundation for complete structure determination of E protein and for structure-based drug discovery targeting this protein.","journal":"PLoS Pathogens","year":2021,"id":164204,"datarank":0.5533319181170905,"base_score":3.6888794541139363,"endowment":3.6888794541139363,"self_citation_contribution":0.5533319181170905,"citation_network_contribution":0.0,"self_endowment_contribution":0.5533319181170905,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":39,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9711,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":685062,"name":"Haley Siddiqi","orcid":"0000-0002-6487-4394","position":1,"is_corresponding":false},{"id":685063,"name":"Daniela V. Castro","orcid":"0000-0002-5423-9821","position":2,"is_corresponding":false},{"id":685064,"name":"Anna A. De Angelis","orcid":"0000-0002-0754-7805","position":3,"is_corresponding":false},{"id":621845,"name":"Aaron L. Oom","orcid":"0000-0003-3300-0333","position":4,"is_corresponding":false},{"id":314249,"name":"Charlotte A. Stoneham","orcid":"0000-0003-3729-6936","position":5,"is_corresponding":false},{"id":621842,"name":"Mary K. Lewinski","orcid":"0000-0002-3784-9660","position":6,"is_corresponding":false},{"id":249772,"name":"Alex E. Clark","orcid":"0000-0003-1687-416X","position":7,"is_corresponding":false},{"id":480576,"name":"Ben A. Croker","orcid":"0000-0002-0885-3599","position":8,"is_corresponding":false},{"id":103778,"name":"Aaron F. Carlin","orcid":"0000-0002-1669-8066","position":9,"is_corresponding":false},{"id":315948,"name":"John Guatelli","orcid":null,"position":10,"is_corresponding":false},{"id":406124,"name":"Stanley J. Opella","orcid":"0000-0003-0443-1396","position":11,"is_corresponding":false},{"id":685061,"name":"Sang Ho Park","orcid":"0000-0003-2973-7156","position":0,"is_corresponding":true}],"reference_count":81,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-18T23:45:27.031465Z","pmid":"34003853","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":[]}