Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning

View ORCID ProfileWenhao O. Ouyang, View ORCID ProfileTimothy J.C. Tan, View ORCID ProfileRuipeng Lei, View ORCID ProfileGe Song, View ORCID ProfileCollin Kieffer, View ORCID ProfileRaiees Andrabi, View ORCID ProfileKenneth A. Matreyek, View ORCID ProfileNicholas C. Wu
doi: https://doi.org/10.1101/2022.06.20.496903
Wenhao O. Ouyang
1Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Wenhao O. Ouyang
Timothy J.C. Tan
2Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Timothy J.C. Tan
Ruipeng Lei
1Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Ruipeng Lei
Ge Song
3Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA
4IAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA 92037, USA
5Consortium for HIV/AIDS Vaccine Development (CHAVD), The Scripps Research Institute, La Jolla, CA 92037, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Ge Song
Collin Kieffer
6Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Collin Kieffer
Raiees Andrabi
3Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA
4IAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA 92037, USA
5Consortium for HIV/AIDS Vaccine Development (CHAVD), The Scripps Research Institute, La Jolla, CA 92037, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Raiees Andrabi
Kenneth A. Matreyek
7Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Kenneth A. Matreyek
Nicholas C. Wu
1Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
2Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
8Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
9Carle Illinois College of Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Nicholas C. Wu
  • For correspondence: nicwu@illinois.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Data/Code
  • Preview PDF
Loading

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.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵* These authors contributed equally to this work

  • https://github.com/nicwulab/SARS-CoV-2_NTD_DMS

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Back to top
PreviousNext
Posted June 21, 2022.
Download PDF

Supplementary Material

Data/Code
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning
Wenhao O. Ouyang, Timothy J.C. Tan, Ruipeng Lei, Ge Song, Collin Kieffer, Raiees Andrabi, Kenneth A. Matreyek, Nicholas C. Wu
bioRxiv 2022.06.20.496903; doi: https://doi.org/10.1101/2022.06.20.496903
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Probing the biophysical constraints of SARS-CoV-2 spike N-terminal domain using deep mutational scanning
Wenhao O. Ouyang, Timothy J.C. Tan, Ruipeng Lei, Ge Song, Collin Kieffer, Raiees Andrabi, Kenneth A. Matreyek, Nicholas C. Wu
bioRxiv 2022.06.20.496903; doi: https://doi.org/10.1101/2022.06.20.496903

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (4667)
  • Biochemistry (10332)
  • Bioengineering (7653)
  • Bioinformatics (26277)
  • Biophysics (13497)
  • Cancer Biology (10663)
  • Cell Biology (15389)
  • Clinical Trials (138)
  • Developmental Biology (8480)
  • Ecology (12800)
  • Epidemiology (2067)
  • Evolutionary Biology (16817)
  • Genetics (11378)
  • Genomics (15451)
  • Immunology (10591)
  • Microbiology (25141)
  • Molecular Biology (10187)
  • Neuroscience (54317)
  • Paleontology (399)
  • Pathology (1663)
  • Pharmacology and Toxicology (2889)
  • Physiology (4331)
  • Plant Biology (9223)
  • Scientific Communication and Education (1585)
  • Synthetic Biology (2551)
  • Systems Biology (6769)
  • Zoology (1459)