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

Abstract

Summary: 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.For complete details on the use and execution of this protocol, please refer to Cecon et al. (2021).

Keywords

• Cell Biology
• Cell culture
• Cell-based Assays
• High Throughput Screening
• Molecular Biology
• Molecular/Chemical Probes