1. A thermostable, closed SARS-CoV-2 spike protein trimer
- Author
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Xiong, Xiaoli, Qu, Kun, Ciazynska, Katarzyna A., Hosmillo, Myra, Carter, Andrew P., Ebrahimi, Soraya, Ke, Zunlong, Scheres, Sjors H. W., Bergamaschi, Laura, Grice, Guinevere L., Zhang, Ying, Bradley, John, Lyons, Paul A., Smith, Kenneth G. C., Toshner, Mark, Elmer, Anne, Ribeiro, Carla, Kourampa, Jenny, Jose, Sherly, Kennet, Jane, Rowlands, Jane, Meadows, Anne, O’Brien, Criona, Rastall, Rebecca, Crucusio, Cherry, Hewitt, Sarah, Price, Jane, Calder, Jo, Canna, Laura, Bucke, Ashlea, Tordesillas, Hugo, Harris, Julie, Ruffolo, Valentina, Domingo, Jason, Graves, Barbara, Butcher, Helen, Caputo, Daniela, Le Gresley, Emma, Dunmore, Benjamin J., Martin, Jennifer, Legchenko, Ekaterina, Treacy, Carmen, Huang, Christopher, Wood, Jennifer, Sutcliffe, Rachel, Hodgson, Josh, Shih, Joy, Graf, Stefan, Tong, Zhen, Mescia, Federica, Tilly, Tobias, O’Donnell, Ciara, Hunter, Kelvin, Pointon, Linda, Pond, Nicole, Wylot, Marta, Jones, Emma, Fawke, Stuart, Bullman, Ben, Turner, Lori, Jarvis, Isobel, Omarjee, Ommar, De Sa, Aloka, Marsden, Joe, Betancourt, Ariana, Perera, Marianne, Epping, Maddie, Richoz, Nathan, Bower, Georgie, Sharma, Rahul, Nice, Francesca, Huhn, Oisin, Stark, Hannah, Walker, Neil, Stirrups, Kathy, Ovington, Nigel, Dewhust, Eleanor, Li, Emily, Papadia, Sofia, Nathan, James A., Baker, Stephen, James, Leo C., Baxendale, Helen E., Goodfellow, Ian, Doffinger, Rainer, Briggs, John A. G., Hosmillo, Myra [0000-0002-3514-7681], Nathan, James [0000-0002-0248-1632], Baker, Stephen [0000-0003-1308-5755], Goodfellow, Ian [0000-0002-9483-510X], and Apollo - University of Cambridge Repository
- Subjects
Models, Molecular ,Immunogen ,Protein Conformation ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Trimer ,Enzyme-Linked Immunosorbent Assay ,Receptor binding site ,medicine.disease_cause ,Protein Engineering ,Article ,03 medical and health sciences ,Betacoronavirus ,0302 clinical medicine ,Protein structure ,COVID-19 Testing ,Models ,Structural Biology ,medicine ,Humans ,Disulfides ,Molecular Biology ,030304 developmental biology ,Cell entry ,0303 health sciences ,Mutation ,Clinical Laboratory Techniques ,Protein Stability ,SARS-CoV-2 ,Cryoelectron Microscopy ,Temperature ,Spike Protein ,Molecular ,Coronavirus Infections ,Flow Cytometry ,Immunoglobulin G ,Protein Multimerization ,Spike Glycoprotein, Coronavirus ,Spike Glycoprotein ,3. Good health ,Coronavirus ,Membrane ,Biophysics ,030217 neurology & neurosurgery - Abstract
The spike (S) protein of SARS-CoV-2 mediates receptor binding and cell entry and is the dominant target of the immune system. S 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 which allow production of thermostable, crosslinked, S protein trimers that are trapped in the closed, pre-fusion, state. We have determined the structures of crosslinked and non-crosslinked proteins, identifying two distinct closed 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.
- Published
- 2020
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