Conformational changes in the Ebola virus membrane fusion machine induced by pH, Ca2+, and receptor binding.
Allosteric Regulation
Calcium
/ metabolism
Ebolavirus
/ chemistry
Fluorescence Resonance Energy Transfer
HEK293 Cells
Humans
Hydrogen-Ion Concentration
Intracellular Signaling Peptides and Proteins
/ metabolism
Membrane Fusion
Niemann-Pick C1 Protein
Polysaccharides
/ metabolism
Protein Binding
Protein Conformation
Protein Domains
Viral Envelope Proteins
/ chemistry
Virus Internalization
Journal
PLoS biology
ISSN: 1545-7885
Titre abrégé: PLoS Biol
Pays: United States
ID NLM: 101183755
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
16
09
2019
accepted:
23
01
2020
revised:
21
02
2020
pubmed:
11
2
2020
medline:
15
5
2020
entrez:
11
2
2020
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 32040508
doi: 10.1371/journal.pbio.3000626
pii: PBIOLOGY-D-19-02721
pmc: PMC7034923
doi:
Substances chimiques
Intracellular Signaling Peptides and Proteins
0
NPC1 protein, human
0
Niemann-Pick C1 Protein
0
Polysaccharides
0
Viral Envelope Proteins
0
envelope glycoprotein, Ebola virus
0
Calcium
SY7Q814VUP
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
e3000626Subventions
Organisme : NIAID NIH HHS
ID : DP2 AI124384
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI148784
Pays : United States
Organisme : NIGMS NIH HHS
ID : R25 GM066567
Pays : United States
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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