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

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 transmem...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sang Ho Park, Haley Siddiqi, Daniela V Castro, Anna A De Angelis, Aaron L Oom, Charlotte A Stoneham, Mary K Lewinski, Alex E Clark, Ben A Croker, Aaron F Carlin, John Guatelli, Stanley J Opella
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/7c6420860ea3475c9217299c1fbe66ce
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7c6420860ea3475c9217299c1fbe66ce
record_format	dspace
spelling	oai:doaj.org-article:7c6420860ea3475c9217299c1fbe66ce2021-12-02T19:59:40ZInteractions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.1553-73661553-737410.1371/journal.ppat.1009519https://doaj.org/article/7c6420860ea3475c9217299c1fbe66ce2021-05-01T00:00:00Zhttps://doi.org/10.1371/journal.ppat.1009519https://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374SARS-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.Sang Ho ParkHaley SiddiqiDaniela V CastroAnna A De AngelisAaron L OomCharlotte A StonehamMary K LewinskiAlex E ClarkBen A CrokerAaron F CarlinJohn GuatelliStanley J OpellaPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 17, Iss 5, p e1009519 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Sang Ho Park Haley Siddiqi Daniela V Castro Anna A De Angelis Aaron L Oom Charlotte A Stoneham Mary K Lewinski Alex E Clark Ben A Croker Aaron F Carlin John Guatelli Stanley J Opella Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
description	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.
format	article
author	Sang Ho Park Haley Siddiqi Daniela V Castro Anna A De Angelis Aaron L Oom Charlotte A Stoneham Mary K Lewinski Alex E Clark Ben A Croker Aaron F Carlin John Guatelli Stanley J Opella
author_facet	Sang Ho Park Haley Siddiqi Daniela V Castro Anna A De Angelis Aaron L Oom Charlotte A Stoneham Mary K Lewinski Alex E Clark Ben A Croker Aaron F Carlin John Guatelli Stanley J Opella
author_sort	Sang Ho Park
title	Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
title_short	Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
title_full	Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
title_fullStr	Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
title_full_unstemmed	Interactions of SARS-CoV-2 envelope protein with amilorides correlate with antiviral activity.
title_sort	interactions of sars-cov-2 envelope protein with amilorides correlate with antiviral activity.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/7c6420860ea3475c9217299c1fbe66ce
work_keys_str_mv	AT sanghopark interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT haleysiddiqi interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT danielavcastro interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT annaadeangelis interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT aaronloom interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT charlotteastoneham interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT maryklewinski interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT alexeclark interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT benacroker interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT aaronfcarlin interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT johnguatelli interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity AT stanleyjopella interactionsofsarscov2envelopeproteinwithamiloridescorrelatewithantiviralactivity
_version_	1718375731344441344

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

Ejemplares similares