Article ; Online: SARS-COV-2 spike binding to ACE2 in living cells monitored by TR-FRET.
Cell chemical biology
2021 Volume 29, Issue 1, Page(s) 74–83.e4
Abstract: ... we developed an RBD-ACE2 binding assay that is based on time-resolved FRET, which reliably monitors ... Targeting the interaction between the SARS-CoV-2 spike protein and human ACE2, its primary ... revealed that the receptor binding domain (RBD) of the spike protein plays a prominent role in ACE2 binding ...
| 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. |
|---|---|
| MeSH term(s) | Angiotensin-Converting Enzyme 2/chemistry ; Cells, Cultured ; Fluorescence Resonance Energy Transfer ; HEK293 Cells ; Humans ; Protein Binding ; Spike Glycoprotein, Coronavirus/chemistry ; Time Factors |
| Chemical Substances | Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2 ; ACE2 protein, human (EC 3.4.17.23) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23) |
| Language | English |
| Publishing date | 2021-07-02 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ISSN | 2451-9448 |
| ISSN (online) | 2451-9448 |
| DOI | 10.1016/j.chembiol.2021.06.008 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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