Multimerization- and glycosylation-dependent receptor binding of SARS-CoV-2 spike proteins
0301 basic medicine
Glycosylation
QH301-705.5
Immunology
EMC OR-01
N-Acetylglucosaminyltransferases
Microbiology
Madin Darby Canine Kidney Cells
Mice
03 medical and health sciences
Dogs
Virology
Chlorocebus aethiops
Genetics
Animals
Humans
Biology (General)
Molecular Biology
Vero Cells
Mesocricetus
SARS-CoV-2
RC581-607
3. Good health
HEK293 Cells
A549 Cells
Spike Glycoprotein, Coronavirus
Parasitology
Angiotensin-Converting Enzyme 2
Immunologic diseases. Allergy
Protein Multimerization
Research Article
Protein Binding
DOI:
10.1371/journal.ppat.1009282
Publication Date:
2021-02-09T02:03:27Z
AUTHORS (13)
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.
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