STAR Protoc . Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET

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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 ¹ , Julie Dam ¹ , Ralf Jockers ¹



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.