FIG 2.

Lpro deISGylation in vitro activity. (A) Recombinant human proISG15 treated with bacterial extracts expressing WT or indicated mutant forms of Lpro. Lpro-dependent cleavage of proISG15 is detected as a visible 2-kDa band shift. (B) Immunoblot of samples treated with bacterial extracts expressing WT or Lpro mutants. Bacterial extracts prepared from IPTG-induced cultures were mixed with SK6 lysates, followed by incubation at 4°C to allow for Lpro-dependent enzymatic digestion of host proteins. Cleavage of eIF4G (220 kDa) generates a fragment of approximately 120 kDa (CP). For each figure, one representative blot is shown out of three independent experiments.