SARS-COV-2 spike binding to ACE2 in living cells monitored by TR-FRET.
Cell Chem Biol
; 29(1): 74-83.e4, 2022 01 20.
Article
in En
| MEDLINE
| ID: mdl-34246414
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fluorescence Resonance Energy Transfer
/
Spike Glycoprotein, Coronavirus
/
Angiotensin-Converting Enzyme 2
Limits:
Humans
Language:
En
Journal:
Cell Chem Biol
Year:
2022
Document type:
Article
Affiliation country:
France
Country of publication:
United States