SARS-COV-2 spike binding to ACE2 in living cells monitored by TR-FRET - Université Paris Cité Access content directly
Journal Articles Cell Chemical Biology Year : 2022

SARS-COV-2 spike binding to ACE2 in living cells monitored by TR-FRET

Erika Cecon
Matilda Burridge
  • Function : Author
Lauren Carter
Julie Dam
Ralf Jockers

Abstract

Targeting the interaction between the SARS-CoV-2 spike protein and human ACE2, its primary cell membrane receptor, is a promising therapeutic strategy to prevent viral entry. Recent in vitro studies revealed that the receptor binding domain (RBD) of the spike protein plays a prominent role in ACE2 binding, yet a simple and quantitative assay for monitoring this interaction in a cellular environment is lacking. Here, we developed an RBD-ACE2 binding assay that is based on time-resolved FRET, which reliably monitors the interaction in a physiologically relevant and cellular context. Because it is modular, the assay can monitor the impact of different cellular components, such as heparan sulfate, lipids, and membrane proteins on the RBD-ACE2 interaction and it can be extended to the full-length spike protein. The assay is HTS compatible and can detect small-molecule competitive and allosteric modulators of the RBD-ACE2 interaction with high relevance for SARS-CoV-2 therapeutics.
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Dates and versions

hal-03685014 , version 1 (07-11-2022)

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Erika Cecon, Matilda Burridge, Longxing Cao, Lauren Carter, Rashmi Ravichandran, et al.. SARS-COV-2 spike binding to ACE2 in living cells monitored by TR-FRET. Cell Chemical Biology, 2022, 29 (1), pp.74-83. ⟨10.1016/j.chembiol.2021.06.008⟩. ⟨hal-03685014⟩
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