Qualitative analysis of polyUb chain hydrolysis by CCHFV L protein and the isolated CCHFV OTU domain.
Purified CCHFV L protein (RdRp) was incubated with either (A) K48polyUb or (B) K63polyUb chains in the presence or absence of OTU-specific inhibitor CC.4. Ub chain hydrolysis was also assessed for the purified OTU domain using (C) K48polyUb chains or (D) K63polyUb chains in the presence or absence of CC.4. Reactions were incubated at 37°C and samples taken at 0, 15 and 30 minutes as indicated. Ub chain lengths following digestion are indicated with arrows. Samples were resolved on a 10% tris-tricine gel and visualized by silver stain.