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

被引：2
|
作者：
Cecon, Erika [1 ]
Dam, Julie [1 ]
Jockers, Ralf [1 ]
机构：
[1] Univ Paris, Inst Cochin, INSERM, CNRS, F-75014 Paris, France
来源：
STAR PROTOCOLS | 2022年 / 3卷 / 01期
关键词：
Cell Biology; Cell culture; Cell-based Assays; High Throughput Screening; Molecular Biology; Molecular/Chemical Probes;
D O I：
10.1016/j.xpro.2021.101024
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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/ACE 2 interaction in physiologically rele-vant cellular contexts and is suitable for high-throughput investigation of inter-fering small-molecule compounds and antibodies.For complete details on the use and execution of this protocol, please refer to Cecon et al. (2021).
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页数：14
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