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Image Search Results
Journal: The Journal of Biological Chemistry
Article Title: Adapted ATPase domain communication overcomes the cytotoxicity of p97 inhibitors
doi: 10.1074/jbc.RA118.004301
Figure Lengend Snippet: 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.
Article Snippet: The 2-fold serial dilutions of p97 proteins (maximum 40 μ m , purified from bacteria) were mixed with 20 n m BODIPY-FL–ATP (ThermoFisher Scientific) or 10
Techniques: Mutagenesis, Binding Assay, Generated, Labeling, Fluorescence