Development of a SARS-CoV-2-derived receptor-binding domain-based ACE2 biosensor

Jung-Soo Suh; Heon-Su Kim; Tae-Jin Kim

doi:10.1016/j.snb.2021.129663

Development of a SARS-CoV-2-derived receptor-binding domain-based ACE2 biosensor

Sens Actuators B Chem. 2021 May 1:334:129663. doi: 10.1016/j.snb.2021.129663. Epub 2021 Feb 16.

Authors

Jung-Soo Suh¹, Heon-Su Kim¹, Tae-Jin Kim^{1

2

3}

Affiliations

¹ Department of Integrated Biological Science, Pusan National University, Pusan 46241, Republic of Korea.
² Department of Biological Sciences, Pusan National University, Pusan 46241, Republic of Korea.
³ Institute of Systems Biology, Pusan National University, Pusan 46241, Republic of Korea.

Abstract

The global outbreak of coronavirus disease and rapid spread of the causative severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represent a significant threat to human health. A key mechanism of human SARS-CoV-2 infection is initiated by the combination of human angiotensin-converting enzyme 2 (hACE2) and the receptor-binding domain (RBD) of the SARS-CoV-2-derived spike glycoprotein. Despite the importance of these protein interactions, there are still insufficient detection methods to observe their activity at the cellular level. Herein, we developed a novel fluorescence resonance energy transfer (FRET)-based hACE2 biosensor to monitor the interaction between hACE2 and SARS-CoV-2 RBD. This biosensor facilitated the visualization of hACE2-RBD activity with high spatiotemporal resolutions at the single-cell level. Further studies revealed that the FRET-based hACE2 biosensors were sensitive to both exogenous and endogenous hACE2 expression, suggesting that they might be safely applied to the early stage of SARS-CoV-2 infection without direct virus use. Therefore, our novel biosensor could potentially help develop drugs that target SARS-CoV-2 by inhibiting hACE2-RBD interaction.

Keywords: ACE2; Biosensor; CQ, chloroquine; FRET; HCQ, hydroxychloroquine; Live-cell imaging; NA, numerical aperture; RBD, receptor-binding domain; RBM, receptor-binding motif; ROI, region of interest; SARS-CoV-2; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; SEM, standard error of the mean; bg, background; hACE2, human angiotensin-converting enzyme 2.