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Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics

The Ebola virus (EBOV) envelope glycoprotein (GP) mediates the fusion of the virion membrane with the membrane of susceptible target cells during infection. While proteolytic cleavage of GP by endosomal cathepsins and binding of the cellular receptor Niemann-Pick C1 protein (NPC1) are essential step...

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Autores principales: Durham, Natasha D., Howard, Angela R., Govindan, Ramesh, Senjobe, Fernando, Fels, J. Maximilian, Diehl, William E., Luban, Jeremy, Chandran, Kartik, Munro, James B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7019799/
https://www.ncbi.nlm.nih.gov/pubmed/31952255
http://dx.doi.org/10.3390/v12010103
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author Durham, Natasha D.
Howard, Angela R.
Govindan, Ramesh
Senjobe, Fernando
Fels, J. Maximilian
Diehl, William E.
Luban, Jeremy
Chandran, Kartik
Munro, James B.
author_facet Durham, Natasha D.
Howard, Angela R.
Govindan, Ramesh
Senjobe, Fernando
Fels, J. Maximilian
Diehl, William E.
Luban, Jeremy
Chandran, Kartik
Munro, James B.
author_sort Durham, Natasha D.
collection PubMed
description The Ebola virus (EBOV) envelope glycoprotein (GP) mediates the fusion of the virion membrane with the membrane of susceptible target cells during infection. While proteolytic cleavage of GP by endosomal cathepsins and binding of the cellular receptor Niemann-Pick C1 protein (NPC1) are essential steps for virus entry, the detailed mechanisms by which these events promote membrane fusion remain unknown. Here, we applied single-molecule Förster resonance energy transfer (smFRET) imaging to investigate the structural dynamics of the EBOV GP trimeric ectodomain, and the functional transmembrane protein on the surface of pseudovirions. We show that in both contexts, pre-fusion GP is dynamic and samples multiple conformations. Removal of the glycan cap and NPC1 binding shift the conformational equilibrium, suggesting stabilization of conformations relevant to viral fusion. Furthermore, several neutralizing antibodies enrich alternative conformational states. This suggests that these antibodies neutralize EBOV by restricting access to GP conformations relevant to fusion. This work demonstrates previously unobserved dynamics of pre-fusion EBOV GP and presents a platform with heightened sensitivity to conformational changes for the study of GP function and antibody-mediated neutralization.
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spelling pubmed-70197992020-03-09 Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics Durham, Natasha D. Howard, Angela R. Govindan, Ramesh Senjobe, Fernando Fels, J. Maximilian Diehl, William E. Luban, Jeremy Chandran, Kartik Munro, James B. Viruses Article The Ebola virus (EBOV) envelope glycoprotein (GP) mediates the fusion of the virion membrane with the membrane of susceptible target cells during infection. While proteolytic cleavage of GP by endosomal cathepsins and binding of the cellular receptor Niemann-Pick C1 protein (NPC1) are essential steps for virus entry, the detailed mechanisms by which these events promote membrane fusion remain unknown. Here, we applied single-molecule Förster resonance energy transfer (smFRET) imaging to investigate the structural dynamics of the EBOV GP trimeric ectodomain, and the functional transmembrane protein on the surface of pseudovirions. We show that in both contexts, pre-fusion GP is dynamic and samples multiple conformations. Removal of the glycan cap and NPC1 binding shift the conformational equilibrium, suggesting stabilization of conformations relevant to viral fusion. Furthermore, several neutralizing antibodies enrich alternative conformational states. This suggests that these antibodies neutralize EBOV by restricting access to GP conformations relevant to fusion. This work demonstrates previously unobserved dynamics of pre-fusion EBOV GP and presents a platform with heightened sensitivity to conformational changes for the study of GP function and antibody-mediated neutralization. MDPI 2020-01-15 /pmc/articles/PMC7019799/ /pubmed/31952255 http://dx.doi.org/10.3390/v12010103 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Durham, Natasha D.
Howard, Angela R.
Govindan, Ramesh
Senjobe, Fernando
Fels, J. Maximilian
Diehl, William E.
Luban, Jeremy
Chandran, Kartik
Munro, James B.
Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title_full Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title_fullStr Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title_full_unstemmed Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title_short Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics
title_sort real-time analysis of individual ebola virus glycoproteins reveals pre-fusion, entry-relevant conformational dynamics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7019799/
https://www.ncbi.nlm.nih.gov/pubmed/31952255
http://dx.doi.org/10.3390/v12010103
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