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Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike

The spike (S) protein of SARS-CoV-2 has been observed in three distinct pre-fusion conformations: locked, closed and open. Of these, the function of the locked conformation remains poorly understood. Here we engineered a SARS-CoV-2 S protein construct “S-R/x3” to arrest SARS-CoV-2 spikes in the lock...

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Autores principales: Qu, Kun, Chen, Qiuluan, Ciazynska, Katarzyna A., Liu, Banghui, Zhang, Xixi, Wang, Jingjing, He, Yujie, Guan, Jiali, He, Jun, Liu, Tian, Zhang, Xiaofei, Carter, Andrew P., Xiong, Xiaoli, Briggs, John A. G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9365160/
https://www.ncbi.nlm.nih.gov/pubmed/35905112
http://dx.doi.org/10.1371/journal.ppat.1010583
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author Qu, Kun
Chen, Qiuluan
Ciazynska, Katarzyna A.
Liu, Banghui
Zhang, Xixi
Wang, Jingjing
He, Yujie
Guan, Jiali
He, Jun
Liu, Tian
Zhang, Xiaofei
Carter, Andrew P.
Xiong, Xiaoli
Briggs, John A. G.
author_facet Qu, Kun
Chen, Qiuluan
Ciazynska, Katarzyna A.
Liu, Banghui
Zhang, Xixi
Wang, Jingjing
He, Yujie
Guan, Jiali
He, Jun
Liu, Tian
Zhang, Xiaofei
Carter, Andrew P.
Xiong, Xiaoli
Briggs, John A. G.
author_sort Qu, Kun
collection PubMed
description The spike (S) protein of SARS-CoV-2 has been observed in three distinct pre-fusion conformations: locked, closed and open. Of these, the function of the locked conformation remains poorly understood. Here we engineered a SARS-CoV-2 S protein construct “S-R/x3” to arrest SARS-CoV-2 spikes in the locked conformation by a disulfide bond. Using this construct we determined high-resolution structures confirming that the x3 disulfide bond has the ability to stabilize the otherwise transient locked conformations. Structural analyses reveal that wild-type SARS-CoV-2 spike can adopt two distinct locked-1 and locked-2 conformations. For the D614G spike, based on which all variants of concern were evolved, only the locked-2 conformation was observed. Analysis of the structures suggests that rigidified domain D in the locked conformations interacts with the hinge to domain C and thereby restrains RBD movement. Structural change in domain D correlates with spike conformational change. We propose that the locked-1 and locked-2 conformations of S are present in the acidic high-lipid cellular compartments during virus assembly and egress. In this model, release of the virion into the neutral pH extracellular space would favour transition to the closed or open conformations. The dynamics of this transition can be altered by mutations that modulate domain D structure, as is the case for the D614G mutation, leading to changes in viral fitness. The S-R/x3 construct provides a tool for the further structural and functional characterization of the locked conformations of S, as well as how sequence changes might alter S assembly and regulation of receptor binding domain dynamics.
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spelling pubmed-93651602022-08-11 Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike Qu, Kun Chen, Qiuluan Ciazynska, Katarzyna A. Liu, Banghui Zhang, Xixi Wang, Jingjing He, Yujie Guan, Jiali He, Jun Liu, Tian Zhang, Xiaofei Carter, Andrew P. Xiong, Xiaoli Briggs, John A. G. PLoS Pathog Research Article The spike (S) protein of SARS-CoV-2 has been observed in three distinct pre-fusion conformations: locked, closed and open. Of these, the function of the locked conformation remains poorly understood. Here we engineered a SARS-CoV-2 S protein construct “S-R/x3” to arrest SARS-CoV-2 spikes in the locked conformation by a disulfide bond. Using this construct we determined high-resolution structures confirming that the x3 disulfide bond has the ability to stabilize the otherwise transient locked conformations. Structural analyses reveal that wild-type SARS-CoV-2 spike can adopt two distinct locked-1 and locked-2 conformations. For the D614G spike, based on which all variants of concern were evolved, only the locked-2 conformation was observed. Analysis of the structures suggests that rigidified domain D in the locked conformations interacts with the hinge to domain C and thereby restrains RBD movement. Structural change in domain D correlates with spike conformational change. We propose that the locked-1 and locked-2 conformations of S are present in the acidic high-lipid cellular compartments during virus assembly and egress. In this model, release of the virion into the neutral pH extracellular space would favour transition to the closed or open conformations. The dynamics of this transition can be altered by mutations that modulate domain D structure, as is the case for the D614G mutation, leading to changes in viral fitness. The S-R/x3 construct provides a tool for the further structural and functional characterization of the locked conformations of S, as well as how sequence changes might alter S assembly and regulation of receptor binding domain dynamics. Public Library of Science 2022-07-29 /pmc/articles/PMC9365160/ /pubmed/35905112 http://dx.doi.org/10.1371/journal.ppat.1010583 Text en © 2022 Qu et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Qu, Kun
Chen, Qiuluan
Ciazynska, Katarzyna A.
Liu, Banghui
Zhang, Xixi
Wang, Jingjing
He, Yujie
Guan, Jiali
He, Jun
Liu, Tian
Zhang, Xiaofei
Carter, Andrew P.
Xiong, Xiaoli
Briggs, John A. G.
Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title_full Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title_fullStr Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title_full_unstemmed Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title_short Engineered disulfide reveals structural dynamics of locked SARS-CoV-2 spike
title_sort engineered disulfide reveals structural dynamics of locked sars-cov-2 spike
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9365160/
https://www.ncbi.nlm.nih.gov/pubmed/35905112
http://dx.doi.org/10.1371/journal.ppat.1010583
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