Fig. 5: PEAK3 homodimerization and both 14-3-3 binding interfaces are critical for PEAK3/14-3-3 complex formation.
From: Structural insights into regulation of the PEAK3 pseudokinase scaffold by 14-3-3
a Diagram of PEAK3 mutants generated to probe PEAK3 binding to the primary site on 14-3-3. b Co-immunoprecipitation of endogenous 14-3-3 with FLAG-tagged WT PEAK3 and PEAK3 mutants (Δ14-3-3, S69A) transiently expressed in HEK293 cells. c Representative co-immunoprecipitation of endogenous 14-3-3 with PEAK3 variants carrying mutations in the secondary 14-3-3 binding interface (R147A, S225A, K293A, W298A, K293A/W298A) transiently expressed in HEK293 cells. d Quantification of co-immunoprecipitation data shown in panel (c) plotted as the mean with standard deviation from 3 independent experiments. Statistical significance was determined using One-way ANOVA Dunnett’s multiple comparisons test, *p < 0.05, **<0.01, ***p < 0.001, ****p < 0.0001. e Co-immunoprecipitation of endogenous 14-3-3 with homodimerization-deficient PEAK3 mutants (L146E, A436E, C453E) transiently expressed in HEK293 cells. f Co-immunoprecipitation of FLAG-tagged and HA-tagged WT PEAK3 and PEAK3 S69A transiently expressed in HEK293 cells. In panels b–f, protein levels were detected with the indicated antibodies by Western Blot. All co-immunoprecipitation data are representative of at least 3 independent experiments. Source data are provided as a Source Data file.
