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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Cecon, Erika, Dam, Julie, Jockers, Ralf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8606263/
https://www.ncbi.nlm.nih.gov/pubmed/34841271
http://dx.doi.org/10.1016/j.xpro.2021.101024
_version_ 1784602308072439808
author Cecon, Erika
Dam, Julie
Jockers, Ralf
author_facet Cecon, Erika
Dam, Julie
Jockers, Ralf
author_sort Cecon, Erika
collection PubMed
description 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).
format Online
Article
Text
id pubmed-8606263
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-86062632021-11-22 Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET Cecon, Erika Dam, Julie Jockers, Ralf STAR Protoc Protocol 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). Elsevier 2021-11-22 /pmc/articles/PMC8606263/ /pubmed/34841271 http://dx.doi.org/10.1016/j.xpro.2021.101024 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Cecon, Erika
Dam, Julie
Jockers, Ralf
Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title_full Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title_fullStr Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title_full_unstemmed Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title_short Detection of SARS-CoV-2 spike protein binding to ACE2 in living cells by TR-FRET
title_sort detection of sars-cov-2 spike protein binding to ace2 in living cells by tr-fret
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8606263/
https://www.ncbi.nlm.nih.gov/pubmed/34841271
http://dx.doi.org/10.1016/j.xpro.2021.101024
work_keys_str_mv AT ceconerika detectionofsarscov2spikeproteinbindingtoace2inlivingcellsbytrfret
AT damjulie detectionofsarscov2spikeproteinbindingtoace2inlivingcellsbytrfret
AT jockersralf detectionofsarscov2spikeproteinbindingtoace2inlivingcellsbytrfret

Ejemplares similares