Abstract
Receptor binding studies on sarbecoviruses would benefit from an available toolkit of recombinant spike proteins, or domains thereof, that recapitulate receptor binding properties of native viruses. We hypothesized that trimeric Receptor Binding Domain (RBD) proteins would be suitable candidates to study receptor binding properties of SARS-CoV-1 and -2. Here we created monomeric and trimeric fluorescent RBD proteins, derived from adherent HEK293T, as well as in GnTI-/- mutant cells, to analyze the effect of complex vs high mannose glycosylation on receptor binding. The results demonstrate that trimeric, complex glycosylated proteins are superior in receptor binding compared to monomeric and immaturely glycosylated variants. Although differences in binding to commonly used cell lines were minimal between the different RBD preparations, substantial differences were observed when respiratory tissues of experimental animals were stained. The RBD trimers demonstrated distinct ACE2 expression profiles in bronchiolar ducts and confirmed the higher binding affinity of SARS-CoV-2 over SARS-CoV-1. Our results show that complex glycosylated trimeric RBD proteins are attractive to analyze sarbecovirus receptor binding and explore ACE2 expression profiles in tissues.
Funder
European Research Council
Koninklijke Nederlandse Akademie van Wetenschappen
Bill and Melinda Gates Foundation
National Institutes of Health
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
Publisher
Public Library of Science (PLoS)
Subject
Virology,Genetics,Molecular Biology,Immunology,Microbiology,Parasitology
Reference47 articles.
1. Human monoclonal antibodies block the binding of SARS-CoV-2 spike protein to angiotensin converting enzyme 2 receptor;X Chen;Cell Mol Immunol,2020
2. Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2;Q Wang;Cell,2020
3. Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses;M Letko;Nature Micro.,2020
4. Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein;AC Walls;Cell,2020
5. A serological assay to detect SARS-CoV-2 seroconversion in humans;F Amanat;Nature Med,2020
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