FIGURE 6.
The NS3hel part does not significantly affect the functionality of the NS2B-NS3pro part. A, kinetics of the Pyr-RTKR-AMC cleavage by NS2B-NS3pro-hel K48A and NS2B-NS3pro K48A. The samples were coincubated with increasing concentrations of the substrate and the Vmax, Km, kcat, and kcat/Km parameters of the cleavage reactions were calculated. B, nucleotides do not affect the proteolytic activity of NS2B-NS3pro-hel K48A and NS2B-NS3pro K48A. The proteolytic activity of the samples (50 nm each) was determined against Pyr-RTKR-AMC in the presence of ssRNA, ssDNA, dsRNA, and DNA dsDNA (200 nm each). The reactions also contained 10 mm ATP. The experiments shown in A and B were repeated three times. The means of triplicate values from a representative experiment are shown.