FIGURE 4.
Dimer-monomer distributions and FRET experiment used for calculation of the dimerization KD of 14-3-3ζ variants. (A) The populations of 14-3-3ζ proteins (x-axis) in the dimeric and monomeric state determined from SEC-RALS experiments (Supplementary Figure S4) as described in the text. The dimer-monomer populations for 164 μM ζ_S58D and ζ_S57D_S58D mutants and 183 μM ζm and pζ variants at 20°C are shown. (B) FRET kinetic profile of ζ_S58E dimerization at 15°C. The experimental data (black dots) were fitted (red line), as in Trošanová et al. (2022), to obtain KD and koff values.