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Inhibition of SARS-CoV-2 PLpro enzyme activities by 9-AMN. The enzymatic inhibitory effects of 9-AMN and minocycline (each at 50 µM) on the SARS-CoV-2 PLpro enzyme were evaluated as detailed in the materials and methods. ( A ) Inhibition of proteolytic activity. ( B ) Inhibition of DUB activity. ( C ) Gradual reduction of proteolytic activity with increasing 9-AMN concentration. ( D ) Gradual decline of DUB activity with increasing 9-AMN concentration. ( E ) Inhibition of 3CLpro enzyme activity ( F ) Inhibition of RdRp enzyme activity. The enzymatic activity of control samples treated with DMSO (0.1%) is taken as 100% for calculating relative enzymatic activity. GRL-0617 (50 µM) treated wells were used as a specificity control. The graph is representative of three independent experiments (n = 3). The P value < 0.05 is denoted as

Journal: Scientific Reports

Article Title: 9-aminominocycline potentiates the efficacy of EIDD-1931 and PF-332 by targeting the papain like protease enzyme of SARS-CoV-2

doi: 10.1038/s41598-025-89717-3

Figure Lengend Snippet: Inhibition of SARS-CoV-2 PLpro enzyme activities by 9-AMN. The enzymatic inhibitory effects of 9-AMN and minocycline (each at 50 µM) on the SARS-CoV-2 PLpro enzyme were evaluated as detailed in the materials and methods. ( A ) Inhibition of proteolytic activity. ( B ) Inhibition of DUB activity. ( C ) Gradual reduction of proteolytic activity with increasing 9-AMN concentration. ( D ) Gradual decline of DUB activity with increasing 9-AMN concentration. ( E ) Inhibition of 3CLpro enzyme activity ( F ) Inhibition of RdRp enzyme activity. The enzymatic activity of control samples treated with DMSO (0.1%) is taken as 100% for calculating relative enzymatic activity. GRL-0617 (50 µM) treated wells were used as a specificity control. The graph is representative of three independent experiments (n = 3). The P value < 0.05 is denoted as "*" indicating statistical significance, while “ns” refers to statistically not significant.

Article Snippet: The recombinant full-length untagged 3CLpro (cat#100823) and His-tagged PLpro (cat#100735), assay buffers (cat#79956 and cat#78039-1 for 3CLpro and PLpro, respectively), GC-367 (cat#78013), and fluorescently labeled substrates (cat#79952 for 3CLpro, cat#79997 for PLpro protease activity, and cat#82200 for DUB activity) were purchased from BPS Biosciences (San Diego, CA, USA).

Techniques: Inhibition, Activity Assay, Concentration Assay, Control