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

被引：200

作者：

Xiong, Xiaoli ^{[1
]}

Qu, Kun ^{[1
]}

Ciazynska, Katarzyna A. ^{[1
]}

Hosmillo, Myra ^{[2
]}

Carter, Andrew P. ^{[1
]}

Ebrahimi, Soraya ^{[3
]}

Ke, Zunlong ^{[1
]}

Scheres, Sjors H. W. ^{[1
]}

Bergamaschi, Laura ^{[4
,9
,11
]}

Grice, Guinevere L. ^{[4
]}

Zhang, Ying ^{[5
,6
]}

Nathan, James A. ^{[4
,9
,10
]}

Baker, Stephen ^{[4
,9
,11
]}

James, Leo C. ^{[7
,9
,11
]}

Baxendale, Helen E. ^{[8
,9
]}

Goodfellow, Ian ^{[2
,12
]}

Doffinger, Rainer ^{[3
,12
]}

Briggs, John A. G. ^{[1
,12
]}

Bradley, John ^{[9
,10
,12
]}

Lyons, Paul A. ^{[9
,11
,13
]}

Smith, Kenneth G. C. ^{[9
,11
,14
]}

Toshner, Mark ^{[9
,14
]}

Elmer, Anne ^{[12
,15
]}

Ribeiro, Carla ^{[12
,14
]}

Kourampa, Jenny ^{[12
,14
]}

Jose, Sherly ^{[12
,14
]}

Kennet, Jane ^{[13
,14
]}

Rowlands, Jane ^{[14
]}

Meadows, Anne ^{[14
]}

O'Brien, Criona ^{[14
,15
]}

Rastall, Rebecca ^{[14
]}

Crucusio, Cherry ^{[14
,15
]}

Hewitt, Sarah ^{[14
]}

Price, Jane ^{[12
,14
]}

Calder, Jo ^{[14
,16
]}

Canna, Laura ^{[14
,16
]}

Bucke, Ashlea ^{[14
,16
]}

Tordesillas, Hugo ^{[14
,16
]}

Harris, Julie ^{[9
,15
]}

Ruffolo, Valentina ^{[9
,14
]}

Domingo, Jason ^{[9
,12
]}

Graves, Barbara ^{[9
,16
]}

Butcher, Helen ^{[9
,16
]}

Caputo, Daniela ^{[9
,16
]}

Le Gresley, Emma ^{[9
,16
]}

Dunmore, Benjamin J. ^{[9
]}

Martin, Jennifer ^{[9
]}

Legchenko, Ekaterina ^{[9
]}

Treacy, Carmen ^{[9
]}

Huang, Christopher ^{[9
,11
]}

机构：

[1] Med Res Council Lab Mol Biol, Struct Studies Div, Cambridge, England

[2] Univ Cambridge, Dept Pathol, Div Virol, Cambridge, England

[3] Addenbrookes Hosp, Dept Clin Biochem & Immunol, Cambridge, England

[4] Univ Cambridge, Cambridge Inst Therapeut Immunol & Infect Dis CIT, Cambridge, England

[5] Sun Yat Sen Univ, Affiliated Hosp 7, Precis Med Ctr, Shenzhen, Guangdong, Peoples R China

[6] Francis Crick Inst, London, England

[7] Med Res Council Lab Mol Biol, Prot & Nucle Acid Chem Div, Cambridge, England

[8] Royal Papworth Hosp NHS Fdn Trust, Cambridge, England

[9] Univ Cambridge, Dept Med, Cambridge, England

[10] Natl Inst Hlth Res Cambridge Biomed Res Ctr, Cambridge, England

[11] Univ Cambridge, Cambridge Inst Therapeut Immunol & Infect Dis, Jeffrey Cheah Biomed Ctr, Cambridge BioMed Campus, Cambridge, England

[12] Natl Inst Hlth Res Clin Res Facil, Cambridge, England

[13] Univ Cambridge, Inst Metab Sci, Metab Res Labs, Cambridge, England

[14] Cambridge Univ Hosp NHS Fdn Trust, Cambridge, England

[15] Univ Cambridge, Dept Paediat, Cambridge, England

[16] Natl Inst Hlth Res BioResource, Cambridge Biomed Campus, Cambridge, England

[17] Univ Cambridge, Dept Psychiat, Cambridge, England

[18] Univ Cambridge, Med Res Council Lab Mol Biol, Mol Immunity Unit, Cambridge, England

[19] Univ Cambridge, Dept Oncol, Cambridge, England

[20] Univ Cambridge, Dept Obstet & Gynaecol, Cambridge, England

来源：

NATURE STRUCTURAL & MOLECULAR BIOLOGY | 2020年 / 27卷 / 10期

基金：

英国医学研究理事会; 欧洲研究理事会; 英国惠康基金;

关键词：

BEAM-INDUCED MOTION; CRYO-EM; CORONAVIRUS; VACCINE;

D O I：

10.1038/s41594-020-0478-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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. The SARS-CoV-2 spike glycoprotein is flexible, and its receptor-binding domain (RBD) fluctuates between open and closed conformations. Disulfide bonds are engineered into the spike ectodomain to lock the RBD in the closed state, leading to a construct with high thermostability.

引用

页码：934 / +

页数：16

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