We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Stabilization of the SARS-CoV-2 receptor binding domain by protein core redesign and deep mutational scanning.
- Authors
Leonard, Alison C; Weinstein, Jonathan J; Steiner, Paul J; Erbse, Annette H; Fleishman, Sarel J; Whitehead, Timothy A
- Abstract
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.
- Subjects
PROTEIN domains; CARRIER proteins; PROTEIN binding; SARS-CoV-2; BINDING sites; MONOCLONAL antibodies
- Publication
PEDS: Protein Engineering, Design & Selection, 2022, Vol 35, p1
- ISSN
1741-0126
- Publication type
Article
- DOI
10.1093/protein/gzac002