Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity

SARS-CoV-2 is the novel coronavirus that causative agent of COVID-19, a sometimes-lethal respiratory infection responsible for world-wide pandemic. The envelope (E) protein, one four structural proteins encoded in viral genome, 75-residue integral membrane protein whose transmembrane domain exhibits ion channel activity and cytoplasmic participates protein-protein interactions. These activities contribute to several aspects replication-cycle, including virion assembly, budding, release, pathogenesis. Here, we describe structure dynamics full-length E hexadecylphosphocholine micelles by NMR spectroscopy. We also characterized its interactions with putative inhibitors. chemical shift index dipolar wave plots establish consists long helix (residues 8–43) short 53–60) connected complex linker some internal mobility. conformations N-terminal C-terminal are unaffected truncation from intact protein. perturbations spectra induced addition inhibitors demonstrate region 6–18) principal binding site. affinity correlates their antiviral potency Vero E6 cells: HMA ? EIPA > DMA >> Amiloride, suggesting bulky hydrophobic groups 5’ position amiloride pyrazine ring play essential roles activity. An N15A mutation increased production virus-like particles, significant changes residues inhibitor site, abolished binding, Asn15 plays key role maintaining conformation near studies provide foundation complete determination structure-based drug discovery targeting this