A Dual‐Color Fluorescent Probe Allows Simultaneous Imaging of Main and Papain‐like Proteases of SARS‐CoV‐2‐Infected Cells for Accurate Detection and Rapid Inhibitor Screening
Angewandte Chemie
; 134(9), 2022.
Article
in English
| ProQuest Central | ID: covidwho-1680266
ABSTRACT
The main protease (Mpro) and papain‐like protease (PLpro) play critical roles in SARS‐CoV‐2 replication and are promising targets for antiviral inhibitors. The simultaneous visualization of Mpro and PLpro is extremely valuable for SARS‐CoV‐2 detection and rapid inhibitor screening. However, such a crucial investigation has remained challenging because of the lack of suitable probes. We have now developed a dual‐color probe (3MBP5) for the simultaneous detection of Mpro and PLpro by fluorescence (or Förster) resonance energy transfer (FRET). This probe produces fluorescence from both the Cy3 and Cy5 fluorophores that are cleaved by Mpro and PLpro. 3MBP5‐activatable specificity was demonstrated with recombinant proteins, inhibitors, plasmid‐transfected HEK 293T cells, and SARS‐CoV‐2‐infected TMPRSS2‐Vero cells. Results from the dual‐color probe first verified the simultaneous detection and intracellular distribution of SARS‐CoV‐2 Mpro and PLpro. This is a powerful tool for the simultaneous detection of different proteases with value for the rapid screening of inhibitors.
Chemistry; Inhibitor Screening; Main Protease; Papain-Like Protease; SARS-CoV-2; Simultaneous Visualization; Papain; Color; Fluorescent indicators; Vero cells; Chemical compounds; Protease; Fluorescence resonance energy transfer; Severe acute respiratory syndrome; Fluorescence; Inhibitors; Energy transfer; Proteinase inhibitors; Severe acute respiratory syndrome coronavirus 2; Fluorophores; Screening; Proteinase
Full text:
Available
Collection:
Databases of international organizations
Database:
ProQuest Central
Type of study:
Diagnostic study
Language:
English
Journal:
Angewandte Chemie
Year:
2022
Document Type:
Article
Similar
MEDLINE
...
LILACS
LIS