Fig. 5: MKP5 residue Y435 mediates MAPK binding.
From: Dynamic and structural insights into allosteric regulation on MKP5 a dual-specificity phosphatase
Full-length GST-tagged MKP5 WT, Y435 mutants, and C408S were transiently transfected with p38α MAPK-HA, JNK1-β-GFP, or ERK2-HA into COS-7 cells for 48 h. Cells were harvested and lysed, followed by affinity purification of GSH Sepharose beads. a GST affinity complexes and whole cell lysates were immunoblotted with anti-HA and anti-GST antibodies. b GST affinity complexes and whole cell lysates were immunoblotted with anti-GFP and anti-GST antibodies. Graphs shown below represent quantitation of (a) p38α MAPK-HA/GST-MKP5 and b JNK1-GFP/GST-MKP5. Data represent the mean ± SEM of three to four to five independent experiments. Statistical significance shown was generated using a two-way ANOVA.
