A thermostable, closed SARS-CoV-2 spike protein trimer

The CITIID-NIHR COVID-19 BioResource Collaboration

doi:10.1038/s41594-020-0478-5

A thermostable, closed SARS-CoV-2 spike protein trimer

The CITIID-NIHR COVID-19 BioResource Collaboration

Research output: Contribution to journal › Article › peer-review

209 Citations (Scopus)

Abstract

The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. It exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor-binding site and, subsequently, from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S-protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S that allow the production of thermostable, disulfide-bonded S-protein trimers that are trapped in the closed, prefusion state. Structures of the disulfide-stabilized and non-disulfide-stabilized proteins reveal distinct closed and locked conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.

Original language	English
Pages (from-to)	934-941
Number of pages	8
Journal	Nature Structural and Molecular Biology
Volume	27
Issue number	10
DOIs	https://doi.org/10.1038/s41594-020-0478-5
Publication status	Published - Oct 1 2020
Externally published	Yes

Bibliographical note

Funding Information:
We thank the staff of the MRC-LMB for generous support during the COVID-19 pandemic lockdown. We thank the staff of the MRC-LMB EM Facility, in particular G. Sharov and G. Cannone, for supporting EM; J. Grimmett and T. Darling for supporting scientific computing; and P. Edwards for supporting cell culture. We thank T. Nakane for advice on image processing, K. Dent for assistance with sample screening, A. Fountain for assistance with primer design, C. Lu for assistance with bioinformatics, F. Coscia for assistance with EM grid preparation and R. Aricescu for advice on protein expression. We thank D. Mallery and D. Paul for performing exploratory experiments. We thank the Royal Papworth Hospital Clinical Staff and R&D team in supporting patient and staff recruitment, J. Gronlund for support with sample collection and Royal Papworth Hospital healthy staff donors for their participation in this research. This study was supported by funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC-CoG-648432 MEMBRANEFUSION to J.A.G.B.), the Medical Research Council as part of United Kingdom Research and Innovation (MC_UP_A025_1011 to A.P.C., MC_UP_A025_1013 to S.H.W.S., MC_U105181010 to L.C.J. and MC_UP_1201/16 to J.A.G.B.), 100 Top Talents Program of Sun Yat-sen University (to Y.Z.) and a Wellcome Trust Senior Fellowship (207498/Z/17/Z to I.G.).

Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.

ASJC Scopus Subject Areas

Structural Biology
Molecular Biology

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1038/s41594-020-0478-5

Cite this

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title = "A thermostable, closed SARS-CoV-2 spike protein trimer",

abstract = "The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. It exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor-binding site and, subsequently, from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S-protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S that allow the production of thermostable, disulfide-bonded S-protein trimers that are trapped in the closed, prefusion state. Structures of the disulfide-stabilized and non-disulfide-stabilized proteins reveal distinct closed and locked conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.",

author = "{The CITIID-NIHR COVID-19 BioResource Collaboration} and Xiaoli Xiong and Kun Qu and Ciazynska, {Katarzyna A.} and Myra Hosmillo and Carter, {Andrew P.} and Soraya Ebrahimi and Zunlong Ke and Scheres, {Sjors H.W.} and Laura Bergamaschi and Grice, {Guinevere L.} and Ying Zhang and John Bradley and Lyons, {Paul A.} and Smith, {Kenneth G.C.} and Mark Toshner and Anne Elmer and Carla Ribeiro and Jenny Kourampa and Sherly Jose and Jane Kennet and Jane Rowlands and Anne Meadows and Criona O{\textquoteright}Brien and Rebecca Rastall and Cherry Crucusio and Sarah Hewitt and Jane Price and Jo Calder and Laura Canna and Ashlea Bucke and Hugo Tordesillas and Julie Harris and Valentina Ruffolo and Jason Domingo and Barbara Graves and Helen Butcher and Daniela Caputo and {Le Gresley}, Emma and Dunmore, {Benjamin J.} and Jennifer Martin and Ekaterina Legchenko and Carmen Treacy and Christopher Huang and Jennifer Wood and Rachel Sutcliffe and Josh Hodgson and Joy Shih and Stefan Graf and Zhen Tong and Federica Mescia",

note = "Funding Information: We thank the staff of the MRC-LMB for generous support during the COVID-19 pandemic lockdown. We thank the staff of the MRC-LMB EM Facility, in particular G. Sharov and G. Cannone, for supporting EM; J. Grimmett and T. Darling for supporting scientific computing; and P. Edwards for supporting cell culture. We thank T. Nakane for advice on image processing, K. Dent for assistance with sample screening, A. Fountain for assistance with primer design, C. Lu for assistance with bioinformatics, F. Coscia for assistance with EM grid preparation and R. Aricescu for advice on protein expression. We thank D. Mallery and D. Paul for performing exploratory experiments. We thank the Royal Papworth Hospital Clinical Staff and R&D team in supporting patient and staff recruitment, J. Gronlund for support with sample collection and Royal Papworth Hospital healthy staff donors for their participation in this research. This study was supported by funding from the European Research Council (ERC) under the European Union{\textquoteright}s Horizon 2020 research and innovation program (ERC-CoG-648432 MEMBRANEFUSION to J.A.G.B.), the Medical Research Council as part of United Kingdom Research and Innovation (MC_UP_A025_1011 to A.P.C., MC_UP_A025_1013 to S.H.W.S., MC_U105181010 to L.C.J. and MC_UP_1201/16 to J.A.G.B.), 100 Top Talents Program of Sun Yat-sen University (to Y.Z.) and a Wellcome Trust Senior Fellowship (207498/Z/17/Z to I.G.). Publisher Copyright: {\textcopyright} 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.",

year = "2020",

month = oct,

day = "1",

doi = "10.1038/s41594-020-0478-5",

language = "English",

volume = "27",

pages = "934--941",

journal = "Nature Structural and Molecular Biology",

issn = "1545-9993",

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T1 - A thermostable, closed SARS-CoV-2 spike protein trimer

AU - The CITIID-NIHR COVID-19 BioResource Collaboration

AU - Xiong, Xiaoli

AU - Qu, Kun

AU - Ciazynska, Katarzyna A.

AU - Hosmillo, Myra

AU - Carter, Andrew P.

AU - Ebrahimi, Soraya

AU - Ke, Zunlong

AU - Scheres, Sjors H.W.

AU - Bergamaschi, Laura

AU - Grice, Guinevere L.

AU - Zhang, Ying

AU - Bradley, John

AU - Lyons, Paul A.

AU - Smith, Kenneth G.C.

AU - Toshner, Mark

AU - Elmer, Anne

AU - Ribeiro, Carla

AU - Kourampa, Jenny

AU - Jose, Sherly

AU - Kennet, Jane

AU - Rowlands, Jane

AU - Meadows, Anne

AU - O’Brien, Criona

AU - Rastall, Rebecca

AU - Crucusio, Cherry

AU - Hewitt, Sarah

AU - Price, Jane

AU - Calder, Jo

AU - Canna, Laura

AU - Bucke, Ashlea

AU - Tordesillas, Hugo

AU - Harris, Julie

AU - Ruffolo, Valentina

AU - Domingo, Jason

AU - Graves, Barbara

AU - Butcher, Helen

AU - Caputo, Daniela

AU - Le Gresley, Emma

AU - Dunmore, Benjamin J.

AU - Martin, Jennifer

AU - Legchenko, Ekaterina

AU - Treacy, Carmen

AU - Huang, Christopher

AU - Wood, Jennifer

AU - Sutcliffe, Rachel

AU - Hodgson, Josh

AU - Shih, Joy

AU - Graf, Stefan

AU - Tong, Zhen

AU - Mescia, Federica

N1 - Funding Information: We thank the staff of the MRC-LMB for generous support during the COVID-19 pandemic lockdown. We thank the staff of the MRC-LMB EM Facility, in particular G. Sharov and G. Cannone, for supporting EM; J. Grimmett and T. Darling for supporting scientific computing; and P. Edwards for supporting cell culture. We thank T. Nakane for advice on image processing, K. Dent for assistance with sample screening, A. Fountain for assistance with primer design, C. Lu for assistance with bioinformatics, F. Coscia for assistance with EM grid preparation and R. Aricescu for advice on protein expression. We thank D. Mallery and D. Paul for performing exploratory experiments. We thank the Royal Papworth Hospital Clinical Staff and R&D team in supporting patient and staff recruitment, J. Gronlund for support with sample collection and Royal Papworth Hospital healthy staff donors for their participation in this research. This study was supported by funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC-CoG-648432 MEMBRANEFUSION to J.A.G.B.), the Medical Research Council as part of United Kingdom Research and Innovation (MC_UP_A025_1011 to A.P.C., MC_UP_A025_1013 to S.H.W.S., MC_U105181010 to L.C.J. and MC_UP_1201/16 to J.A.G.B.), 100 Top Talents Program of Sun Yat-sen University (to Y.Z.) and a Wellcome Trust Senior Fellowship (207498/Z/17/Z to I.G.). Publisher Copyright: © 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.

PY - 2020/10/1

Y1 - 2020/10/1

N2 - The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. It exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor-binding site and, subsequently, from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S-protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S that allow the production of thermostable, disulfide-bonded S-protein trimers that are trapped in the closed, prefusion state. Structures of the disulfide-stabilized and non-disulfide-stabilized proteins reveal distinct closed and locked conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.

AB - The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. It exhibits substantial conformational flexibility. It transitions from closed to open conformations to expose its receptor-binding site and, subsequently, from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. S-protein derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. Here we have designed mutations in S that allow the production of thermostable, disulfide-bonded S-protein trimers that are trapped in the closed, prefusion state. Structures of the disulfide-stabilized and non-disulfide-stabilized proteins reveal distinct closed and locked conformations of the S trimer. We demonstrate that the designed, thermostable, closed S trimer can be used in serological assays. This protein has potential applications as a reagent for serology, virology and as an immunogen.

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A thermostable, closed SARS-CoV-2 spike protein trimer

Abstract

Bibliographical note

ASJC Scopus Subject Areas

PubMed: MeSH publication types

UN SDGs

Access to Document

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