Figure 3.
P472L mutation does not disrupt the binding sites of p97 inhibitors. A, ITC was used to measure the binding of CB-5083 to p97 and the P472L mutant. Top panels are raw data, and lower panels show isotherms with fitted curves and representative KD values. B, effect of increasing NMS-873 concentrations on TR-FRET generated by p97 and the P472L mutant in the presence of BODIPY-FL–ATP, and a terbium-labeled anti-His tag antibody was measured (n = 4, S.D.). Apparent KD values extrapolated from NMS-873–dependent increases in TR-FRET are shown. C, fluorescence polarization experiments were performed (n = 2, S.D.) to measure the enhanced binding of EDA–ADP–ATTO-495 to p97 and the P472L mutant with increasing NMS-873 concentrations. Apparent KD values with S.E. are shown.