tetano
Editor, Senior Moderator
STAR Protoc
. 2021 Nov 22;101024.
doi: 10.1016/j.xpro.2021.101024. Online ahead of print.
Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
Erika Cecon[SUP] 1 [/SUP], Julie Dam[SUP] 1 [/SUP], Ralf Jockers[SUP] 1 [/SUP]
Affiliations
Abstract
The SARS-CoV-2 coronavirus infects human cells through the interaction of the viral envelope Spike protein (IPR044366) with the human Angiotensin-Converting Enzyme 2 (ACE2), expressed at the surface of target cells. Here, we describe a detailed protocol to measure the binding of the receptor binding domain (RBD) of Spike to ACE2 by time-resolved Fluorescence Resonance Energy Transfer (TR-FRET). The assay detects the Spike/ACE2 interaction in physiologically relevant cellular contexts and is suitable for high-throughput investigation of interfering small-molecule compounds and antibodies.
. 2021 Nov 22;101024.
doi: 10.1016/j.xpro.2021.101024. Online ahead of print.
Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
Erika Cecon[SUP] 1 [/SUP], Julie Dam[SUP] 1 [/SUP], Ralf Jockers[SUP] 1 [/SUP]
Affiliations
- PMID: 34841271
- PMCID: PMC8606263
- DOI: 10.1016/j.xpro.2021.101024
Abstract
The SARS-CoV-2 coronavirus infects human cells through the interaction of the viral envelope Spike protein (IPR044366) with the human Angiotensin-Converting Enzyme 2 (ACE2), expressed at the surface of target cells. Here, we describe a detailed protocol to measure the binding of the receptor binding domain (RBD) of Spike to ACE2 by time-resolved Fluorescence Resonance Energy Transfer (TR-FRET). The assay detects the Spike/ACE2 interaction in physiologically relevant cellular contexts and is suitable for high-throughput investigation of interfering small-molecule compounds and antibodies.