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

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

SUBMITTER: Cecon E

PROVIDER: S-EPMC8606263 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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

Cecon Erika E Dam Julie J Jockers Ralf R

STAR protocols 20211122 1

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 suit ...[more]

PMID: 34841271

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