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Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain

Measles virus (MeV) and canine distemper virus (CDV), two members of the Morbillivirus genus, are still causing important global diseases of humans and animals, respectively. To enter target cells, morbilliviruses rely on an envelope-anchored machinery, which is composed of two interacting glycoprot...

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Autores principales: Kalbermatter, David, Shrestha, Neeta, Gall, Flavio M., Wyss, Marianne, Riedl, Rainer, Plattet, Philippe, Fotiadis, Dimitrios
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337061/
https://www.ncbi.nlm.nih.gov/pubmed/32647825
http://dx.doi.org/10.1016/j.yjsbx.2020.100021
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author Kalbermatter, David
Shrestha, Neeta
Gall, Flavio M.
Wyss, Marianne
Riedl, Rainer
Plattet, Philippe
Fotiadis, Dimitrios
author_facet Kalbermatter, David
Shrestha, Neeta
Gall, Flavio M.
Wyss, Marianne
Riedl, Rainer
Plattet, Philippe
Fotiadis, Dimitrios
author_sort Kalbermatter, David
collection PubMed
description Measles virus (MeV) and canine distemper virus (CDV), two members of the Morbillivirus genus, are still causing important global diseases of humans and animals, respectively. To enter target cells, morbilliviruses rely on an envelope-anchored machinery, which is composed of two interacting glycoproteins: a tetrameric receptor binding (H) protein and a trimeric fusion (F) protein. To execute membrane fusion, the F protein initially adopts a metastable, prefusion state that refolds into a highly stable postfusion conformation as the result of a finely coordinated activation process mediated by the H protein. Here, we employed cryo-electron microscopy (cryo-EM) and single particle reconstruction to elucidate the structure of the prefusion state of the CDV F protein ectodomain (solF) at 4.3 Å resolution. Stabilization of the prefusion solF trimer was achieved by fusing the GCNt trimerization sequence at the C-terminal protein region, and expressing and purifying the recombinant protein in the presence of a morbilliviral fusion inhibitor class compound. The three-dimensional cryo-EM map of prefusion CDV solF in complex with the inhibitor clearly shows density for the ligand at the protein binding site suggesting common mechanisms of membrane fusion activation and inhibition employed by different morbillivirus members.
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spelling pubmed-73370612020-07-08 Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain Kalbermatter, David Shrestha, Neeta Gall, Flavio M. Wyss, Marianne Riedl, Rainer Plattet, Philippe Fotiadis, Dimitrios J Struct Biol X Article Measles virus (MeV) and canine distemper virus (CDV), two members of the Morbillivirus genus, are still causing important global diseases of humans and animals, respectively. To enter target cells, morbilliviruses rely on an envelope-anchored machinery, which is composed of two interacting glycoproteins: a tetrameric receptor binding (H) protein and a trimeric fusion (F) protein. To execute membrane fusion, the F protein initially adopts a metastable, prefusion state that refolds into a highly stable postfusion conformation as the result of a finely coordinated activation process mediated by the H protein. Here, we employed cryo-electron microscopy (cryo-EM) and single particle reconstruction to elucidate the structure of the prefusion state of the CDV F protein ectodomain (solF) at 4.3 Å resolution. Stabilization of the prefusion solF trimer was achieved by fusing the GCNt trimerization sequence at the C-terminal protein region, and expressing and purifying the recombinant protein in the presence of a morbilliviral fusion inhibitor class compound. The three-dimensional cryo-EM map of prefusion CDV solF in complex with the inhibitor clearly shows density for the ligand at the protein binding site suggesting common mechanisms of membrane fusion activation and inhibition employed by different morbillivirus members. Elsevier 2020-02-29 /pmc/articles/PMC7337061/ /pubmed/32647825 http://dx.doi.org/10.1016/j.yjsbx.2020.100021 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kalbermatter, David
Shrestha, Neeta
Gall, Flavio M.
Wyss, Marianne
Riedl, Rainer
Plattet, Philippe
Fotiadis, Dimitrios
Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title_full Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title_fullStr Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title_full_unstemmed Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title_short Cryo-EM structure of the prefusion state of canine distemper virus fusion protein ectodomain
title_sort cryo-em structure of the prefusion state of canine distemper virus fusion protein ectodomain
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7337061/
https://www.ncbi.nlm.nih.gov/pubmed/32647825
http://dx.doi.org/10.1016/j.yjsbx.2020.100021
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