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ATP binding to NS3 is antagonized by RNA. NS3 (2 µM) was incubated in the presence or absence of ds9-C20 (20 µM) for 10 min followed by addition of ATP (0–30 mM). Binding was measured by monitoring the change in relative fluorescence of NS3. The excitation wavelength was 295 nm and the emission wavelength was 333 nm. The data were fit to a hyperbola resulting in the Kd value of 0.9 ± 0.1 mM in the absence of RNA and 7 ± 1 mM in the presence of RNA.
