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

Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity

View ORCID ProfileIrene Lui, View ORCID ProfileXin X. Zhou, View ORCID ProfileShion A. Lim, Susanna K. Elledge, Paige Solomon, View ORCID ProfileNicholas J. Rettko, Beth Shoshana Zha, View ORCID ProfileLisa L. Kirkemo, View ORCID ProfileJosef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, View ORCID ProfileKevin K. Leung, James A. Wells
doi: https://doi.org/10.1101/2020.05.21.109157
Irene Lui
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Irene Lui
Xin X. Zhou
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Xin X. Zhou
Shion A. Lim
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Shion A. Lim
Susanna K. Elledge
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paige Solomon
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicholas J. Rettko
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Nicholas J. Rettko
Beth Shoshana Zha
2Department of Pulmonary, Critical Care, Allergy and Sleep Medicine, University of California San Francisco, San Francisco, CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lisa L. Kirkemo
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Lisa L. Kirkemo
Josef A. Gramespacher
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Josef A. Gramespacher
Jia Liu
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Frauke Muecksch
3Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Julio Cesar Cetrulo Lorenzi
4Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Fabian Schmidt
3Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yiska Weisblum
3Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Davide F. Robbiani
4Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michel C. Nussenzweig
4Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA
5Howard Hughes Medical Institute, New York, NY 10016, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Theodora Hatziioannou
3Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paul D. Bieniasz
3Laboratory of Retrovirology, The Rockefeller University, New York, NY 10065, USA
5Howard Hughes Medical Institute, New York, NY 10016, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Oren S. Rosenburg
6Department of Medicine, University of California San Francisco, San Francisco, California, 94158, USA
7Chan Zuckerberg Biohub, San Francisco, CA, 94158, CA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kevin K. Leung
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Kevin K. Leung
James A. Wells
1Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, 94158, USA
7Chan Zuckerberg Biohub, San Francisco, CA, 94158, CA
8Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California, 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: jim.wells@ucsf.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

Abstract

A serious public health crisis is currently unfolding due to the SARS-CoV-2 pandemic. SARS-CoV-2 viral entry depends on an interaction between the receptor binding domain of the trimeric viral Spike protein (Spike-RBD) and the dimeric human angiotensin converting enzyme 2 (ACE2) receptor. While it is clear that strategies to block the Spike/ACE2 interaction are promising as anti-SARS-CoV-2 therapeutics, our current understanding is insufficient for the rational design of maximally effective therapeutic molecules. Here, we investigated the mechanism of Spike/ACE2 interaction by characterizing the binding affinity and kinetics of different multimeric forms of recombinant ACE2 and Spike-RBD domain. We also engineered ACE2 into a split Nanoluciferase-based reporter system to probe the conformational landscape of Spike-RBDs in the context of the Spike trimer. Interestingly, a dimeric form of ACE2, but not monomeric ACE2, binds with high affinity to Spike and blocks viral entry in pseudotyped virus and live SARS-CoV-2 virus neutralization assays. We show that dimeric ACE2 interacts with an RBD on Spike with limited intra-Spike avidity, which nonetheless contributes to the affinity of this interaction. Additionally, we demonstrate that a proportion of Spike can simultaneously interact with multiple ACE2 dimers, indicating that more than one RBD domain in a Spike trimer can adopt an ACE2-accessible “up” conformation. Our findings have significant implications on the design strategies of therapeutic molecules that block the Spike/ACE2 interaction. The constructs we describe are freely available to the research community as molecular tools to further our understanding of SARS-CoV-2 biology.

Competing Interest Statement

The authors have declared no competing interest.

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-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted May 21, 2020.
Download PDF
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.
Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity
(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
Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity
Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung, James A. Wells
bioRxiv 2020.05.21.109157; doi: https://doi.org/10.1101/2020.05.21.109157
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Trimeric SARS-CoV-2 Spike interacts with dimeric ACE2 with limited intra-Spike avidity
Irene Lui, Xin X. Zhou, Shion A. Lim, Susanna K. Elledge, Paige Solomon, Nicholas J. Rettko, Beth Shoshana Zha, Lisa L. Kirkemo, Josef A. Gramespacher, Jia Liu, Frauke Muecksch, Julio Cesar Cetrulo Lorenzi, Fabian Schmidt, Yiska Weisblum, Davide F. Robbiani, Michel C. Nussenzweig, Theodora Hatziioannou, Paul D. Bieniasz, Oren S. Rosenburg, Kevin K. Leung, James A. Wells
bioRxiv 2020.05.21.109157; doi: https://doi.org/10.1101/2020.05.21.109157

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 (4384)
  • Biochemistry (9610)
  • Bioengineering (7104)
  • Bioinformatics (24898)
  • Biophysics (12632)
  • Cancer Biology (9974)
  • Cell Biology (14373)
  • Clinical Trials (138)
  • Developmental Biology (7966)
  • Ecology (12126)
  • Epidemiology (2067)
  • Evolutionary Biology (16003)
  • Genetics (10936)
  • Genomics (14756)
  • Immunology (9881)
  • Microbiology (23699)
  • Molecular Biology (9490)
  • Neuroscience (50926)
  • Paleontology (370)
  • Pathology (1541)
  • Pharmacology and Toxicology (2687)
  • Physiology (4023)
  • Plant Biology (8674)
  • Scientific Communication and Education (1512)
  • Synthetic Biology (2402)
  • Systems Biology (6444)
  • Zoology (1346)