Stabilization of the SARS-CoV-2 receptor binding domain by protein core redesign and deep mutational scanning
© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissionsoup.com..
Stabilizing antigenic proteins as vaccine immunogens or diagnostic reagents is a stringent case of protein engineering and design as the exterior surface must maintain recognition by receptor(s) and antigen-specific antibodies at multiple distinct epitopes. This is a challenge, as stability enhancing mutations must be focused on the protein core, whereas successful computational stabilization algorithms typically select mutations at solvent-facing positions. In this study, we report the stabilization of SARS-CoV-2 Wuhan Hu-1 Spike receptor binding domain using a combination of deep mutational scanning and computational design, including the FuncLib algorithm. Our most successful design encodes I358F, Y365W, T430I, and I513L receptor binding domain mutations, maintains recognition by the receptor ACE2 and a panel of different anti-receptor binding domain monoclonal antibodies, is between 1 and 2°C more thermally stable than the original receptor binding domain using a thermal shift assay, and is less proteolytically sensitive to chymotrypsin and thermolysin than the original receptor binding domain. Our approach could be applied to the computational stabilization of a wide range of proteins without requiring detailed knowledge of active sites or binding epitopes. We envision that this strategy may be particularly powerful for cases when there are multiple or unknown binding sites.
| Errataetall: | |
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:35 |
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| Enthalten in: |
Protein engineering, design & selection : PEDS - 35(2022) vom: 17. Feb. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Leonard, Alison C [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 29.03.2022 Date Revised 25.03.2023 published: Print UpdateOf: bioRxiv. 2021 Nov 24;:. - PMID 34845448 Citation Status MEDLINE |
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| doi: |
10.1093/protein/gzac002 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM338555250 |
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| 245 | 1 | 0 | |a Stabilization of the SARS-CoV-2 receptor binding domain by protein core redesign and deep mutational scanning |
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| 500 | |a Date Revised 25.03.2023 | ||
| 500 | |a published: Print | ||
| 500 | |a UpdateOf: bioRxiv. 2021 Nov 24;:. - PMID 34845448 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissionsoup.com. | ||
| 520 | |a Stabilizing antigenic proteins as vaccine immunogens or diagnostic reagents is a stringent case of protein engineering and design as the exterior surface must maintain recognition by receptor(s) and antigen-specific antibodies at multiple distinct epitopes. This is a challenge, as stability enhancing mutations must be focused on the protein core, whereas successful computational stabilization algorithms typically select mutations at solvent-facing positions. In this study, we report the stabilization of SARS-CoV-2 Wuhan Hu-1 Spike receptor binding domain using a combination of deep mutational scanning and computational design, including the FuncLib algorithm. Our most successful design encodes I358F, Y365W, T430I, and I513L receptor binding domain mutations, maintains recognition by the receptor ACE2 and a panel of different anti-receptor binding domain monoclonal antibodies, is between 1 and 2°C more thermally stable than the original receptor binding domain using a thermal shift assay, and is less proteolytically sensitive to chymotrypsin and thermolysin than the original receptor binding domain. Our approach could be applied to the computational stabilization of a wide range of proteins without requiring detailed knowledge of active sites or binding epitopes. We envision that this strategy may be particularly powerful for cases when there are multiple or unknown binding sites | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a FuncLib | |
| 650 | 4 | |a Rosetta | |
| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 4 | |a deep mutational scanning | |
| 650 | 4 | |a immunogen design | |
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| 700 | 1 | |a Weinstein, Jonathan J |e verfasserin |4 aut | |
| 700 | 1 | |a Steiner, Paul J |e verfasserin |4 aut | |
| 700 | 1 | |a Erbse, Annette H |e verfasserin |4 aut | |
| 700 | 1 | |a Fleishman, Sarel J |e verfasserin |4 aut | |
| 700 | 1 | |a Whitehead, Timothy A |e verfasserin |4 aut | |
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