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. 2021 Jan 17;10(1):66–80. doi: 10.1080/22221751.2020.1865840

Figure 5.

Figure 5.

In vitro activity assays of PDCoV Macro-Ubl2-PLpro and truncated protein. (A) PDCoV Macro-Ubl2-PLpro and truncated constructs. (B) The hydrolytic activity assay was performed with 2 µM enzyme and 40 µM FRET peptides. The Macro-Ubl2-PLpro fluorescence intensity value was set to 100%. (C) The deubiquitinating activity assay was determined with 1 µM enzyme and 0.4 µM Ub-AMC. (D) The deISGylating activity assay was determined with 1 µM enzyme and 0.4 µM ISG15-AMC. Experiments were performed in triplicate, the wild-type fluorescence intensity value was set to 100%. The error bars represent the standard deviations for a minimum of triplicate samples. Asterisks indicate statistical significance calculated using unpaired two-tailed student’s t-test, and values of 0.05 were considered statistically significant. *p < 0.05; **p < 0.01; ***p < 0.001.