Skip to main content
. 2021 Jul 17;297(2):100975. doi: 10.1016/j.jbc.2021.100975

Figure 3.

Figure 3

Fluorescence polarization binding isotherms showing NEDD4L WW domains interacting with individual AMOT PPxY motifs. A–D, binding isotherms of synthetic AMOT PPxY peptides to the four different recombinant NEDD4L WW domains. Binding was measured by changes in the fluorescence polarization (FP) of PPxY peptides with C-terminal Oregon Green fluorophores (pH 7.5 and 25 °C). Color coding: PPxY1 (black), PPxY2 (pink), PPxY3 (green), and the control peptide lacking a PPxY motif (purple). Note that the FP binding isotherm for AMOT1-300 and full-length NEDD4L (violet) is also shown in panel C. Data points report the mean of three independent titrations, and fitted curves follow the equation Y=(maxFP∗[protein]/(Kd + [protein])). Error bars denote SD. E, fitted Kd mean values (μM) ± mean SEM. PPxY, Pro-Pro-x (any amino acid)-Tyr.