Conformational changes in the Ebola virus membrane fusion machine induced by pH, Ca2+, and receptor binding
The Ebola virus (EBOV) envelope glycoprotein (GP) is a membrane fusion machine required for virus entry into cells. Following endocytosis of EBOV, the GP1 domain is cleaved by cellular cathepsins in acidic endosomes, removing the glycan cap and exposing a binding site for the Niemann-Pick C1 (NPC1) receptor. NPC1 binding to cleaved GP1 is required for entry. How this interaction translates to GP2 domain-mediated fusion of viral and endosomal membranes is not known. Here, using a bulk fluorescence dequenching assay and single-molecule Förster resonance energy transfer (smFRET)-imaging, we found that acidic pH, Ca2+, and NPC1 binding synergistically induce conformational changes in GP2 and permit virus-liposome lipid mixing. Acidic pH and Ca2+ shifted the GP2 conformational equilibrium in favor of an intermediate state primed for NPC1 binding. Glycan cap cleavage on GP1 enabled GP2 to transition from a reversible intermediate to an irreversible conformation, suggestive of the postfusion 6-helix bundle; NPC1 binding further promoted transition to the irreversible conformation. Thus, the glycan cap of GP1 may allosterically protect against inactivation of EBOV by premature triggering of GP2.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:18 |
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| Enthalten in: |
PLoS biology - 18(2020), 2 vom: 10. Feb., Seite e3000626 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Das, Dibyendu Kumar [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 14.05.2020 Date Revised 16.04.2022 published: Electronic-eCollection Citation Status MEDLINE |
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| doi: |
10.1371/journal.pbio.3000626 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM30635036X |
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| 100 | 1 | |a Das, Dibyendu Kumar |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Conformational changes in the Ebola virus membrane fusion machine induced by pH, Ca2+, and receptor binding |
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| 500 | |a Date Revised 16.04.2022 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a The Ebola virus (EBOV) envelope glycoprotein (GP) is a membrane fusion machine required for virus entry into cells. Following endocytosis of EBOV, the GP1 domain is cleaved by cellular cathepsins in acidic endosomes, removing the glycan cap and exposing a binding site for the Niemann-Pick C1 (NPC1) receptor. NPC1 binding to cleaved GP1 is required for entry. How this interaction translates to GP2 domain-mediated fusion of viral and endosomal membranes is not known. Here, using a bulk fluorescence dequenching assay and single-molecule Förster resonance energy transfer (smFRET)-imaging, we found that acidic pH, Ca2+, and NPC1 binding synergistically induce conformational changes in GP2 and permit virus-liposome lipid mixing. Acidic pH and Ca2+ shifted the GP2 conformational equilibrium in favor of an intermediate state primed for NPC1 binding. Glycan cap cleavage on GP1 enabled GP2 to transition from a reversible intermediate to an irreversible conformation, suggestive of the postfusion 6-helix bundle; NPC1 binding further promoted transition to the irreversible conformation. Thus, the glycan cap of GP1 may allosterically protect against inactivation of EBOV by premature triggering of GP2 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 7 | |a Intracellular Signaling Peptides and Proteins |2 NLM | |
| 650 | 7 | |a NPC1 protein, human |2 NLM | |
| 650 | 7 | |a Niemann-Pick C1 Protein |2 NLM | |
| 650 | 7 | |a Polysaccharides |2 NLM | |
| 650 | 7 | |a Viral Envelope Proteins |2 NLM | |
| 650 | 7 | |a envelope glycoprotein, Ebola virus |2 NLM | |
| 650 | 7 | |a Calcium |2 NLM | |
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| 700 | 1 | |a Bulow, Uriel |e verfasserin |4 aut | |
| 700 | 1 | |a Diehl, William E |e verfasserin |4 aut | |
| 700 | 1 | |a Durham, Natasha D |e verfasserin |4 aut | |
| 700 | 1 | |a Senjobe, Fernando |e verfasserin |4 aut | |
| 700 | 1 | |a Chandran, Kartik |e verfasserin |4 aut | |
| 700 | 1 | |a Luban, Jeremy |e verfasserin |4 aut | |
| 700 | 1 | |a Munro, James B |e verfasserin |4 aut | |
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