Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning
Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning
ABSTRACT Increasing the expression level of the SARS-CoV-2 spike (S) protein has been critical for COVID-19 vaccine development. While previous efforts largely focused on engineering the receptor-binding domain (RBD) and the S2 subunit, the N-terminal domain (NTD) has been long overlooked due to the limited understanding of its biophysical constraints. In this study, the effects of thousands of NTD single mutations on S protein expression were quantified by deep mutational scanning. Our results revealed that in terms of S protein expression, the mutational tolerability of NTD residues was inversely correlated with their proximity to the RBD and S2. We also identified NTD mutations at the interdomain interface that increased S protein expression without altering its antigenicity. Overall, this study not only advances the understanding of the biophysical constraints of the NTD, but also provides invaluable insights into S-based immunogen design.
Ouyang Wenhao O.、Tan Timothy J.C.、Andrabi Raiees、Song Ge、Wu Nicholas C.、Kieffer Collin、Matreyek Kenneth A.、Lei Ruipeng
Department of Biochemistry, University of Illinois at Urbana-ChampaignCenter for Biophysics and Quantitative Biology, University of Illinois at Urbana-ChampaignDepartment of Immunology and Microbiology, The Scripps Research Institute||IAVI Neutralizing Antibody Center, The Scripps Research Institute||Consortium for HIV/AIDS Vaccine Development (CHAVD), The Scripps Research InstituteDepartment of Immunology and Microbiology, The Scripps Research Institute||IAVI Neutralizing Antibody Center, The Scripps Research Institute||Consortium for HIV/AIDS Vaccine Development (CHAVD), The Scripps Research InstituteDepartment of Biochemistry, University of Illinois at Urbana-Champaign||Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign||Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign||Carle Illinois College of Medicine, University of Illinois at Urbana-ChampaignDepartment of Microbiology, University of Illinois at Urbana-ChampaignDepartment of Pathology, Case Western Reserve University School of MedicineDepartment of Biochemistry, University of Illinois at Urbana-Champaign
生物科学研究方法、生物科学研究技术生物物理学基础医学
Ouyang Wenhao O.,Tan Timothy J.C.,Andrabi Raiees,Song Ge,Wu Nicholas C.,Kieffer Collin,Matreyek Kenneth A.,Lei Ruipeng.Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning[EB/OL].(2025-03-28)[2025-05-12].https://www.biorxiv.org/content/10.1101/2022.06.20.496903.点此复制
评论