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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-01-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/12/1/103 |
| _version_ | 1828855152146120704 |
|---|---|
| author | Natasha D. Durham Angela R. Howard Ramesh Govindan Fernando Senjobe J. Maximilian Fels William E. Diehl Jeremy Luban Kartik Chandran James B. Munro |
| author_facet | Natasha D. Durham Angela R. Howard Ramesh Govindan Fernando Senjobe J. Maximilian Fels William E. Diehl Jeremy Luban Kartik Chandran James B. Munro |
| author_sort | Natasha D. Durham |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-13T00:49:13Z |
| format | Article |
| id | doaj.art-bcdb787ba87e49aa8b5f334b67dd47fe |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-13T00:49:13Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-bcdb787ba87e49aa8b5f334b67dd47fe2022-12-22T00:04:58ZengMDPI AGViruses1999-49152020-01-0112110310.3390/v12010103v12010103Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational DynamicsNatasha D. Durham0Angela R. Howard1Ramesh Govindan2Fernando Senjobe3J. Maximilian Fels4William E. Diehl5Jeremy Luban6Kartik Chandran7James B. Munro8Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USADepartment of Molecular Biology and Microbiology, Tufts University School of Medicine and Sackler School of Graduate Biomedical Sciences, Boston, MA 02111, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USADepartment of Molecular Biology and Microbiology, Tufts University School of Medicine and Sackler School of Graduate Biomedical Sciences, Boston, MA 02111, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USAProgram in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USAProgram in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USADepartment of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USAThe 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.https://www.mdpi.com/1999-4915/12/1/103ebola virusenvelope glycoproteinconformational dynamicssingle-molecule fretvirus entry |
| spellingShingle | Natasha D. Durham Angela R. Howard Ramesh Govindan Fernando Senjobe J. Maximilian Fels William E. Diehl Jeremy Luban Kartik Chandran James B. Munro Real-Time Analysis of Individual Ebola Virus Glycoproteins Reveals Pre-Fusion, Entry-Relevant Conformational Dynamics Viruses ebola virus envelope glycoprotein conformational dynamics single-molecule fret virus entry |
| 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 | ebola virus envelope glycoprotein conformational dynamics single-molecule fret virus entry |
| url | https://www.mdpi.com/1999-4915/12/1/103 |
| work_keys_str_mv | AT natashaddurham realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT angelarhoward realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT rameshgovindan realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT fernandosenjobe realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT jmaximilianfels realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT williamediehl realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT jeremyluban realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT kartikchandran realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics AT jamesbmunro realtimeanalysisofindividualebolavirusglycoproteinsrevealsprefusionentryrelevantconformationaldynamics |