Figure 3.
ITC analysis of PKR binding to VAI. Measurements were performing using a VP-ITC (Microcal, Inc.) at 20°C. Both RNA and protein were exchanged in AU 200 buffer by exhaustive dialysis. The calorimeter cell contained 4 µM VAI and the syringe contained 75 µM PKR. A single 2-µL injection was performed followed by 29 10-µL injections. The top panel shows the background-corrected ITC data and the bottom panel shows the integrated data (points) and fit of the data to a model of two independent binding sites (solid line) performed using ORIGIN (Microcal, Inc.) The best fit parameters are shown in Table 2.