Figure 4: Mutational analysis on interactions between MKP7-CD and JNK1. | Nature Communications

Figure 4: Mutational analysis on interactions between MKP7-CD and JNK1.

From: A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation

Figure 4

(a) Effects of mutations in MKP7-CD on the JNK1 dephosphorylation (mean±s.e.m., n=3). Residues involved in hydrophobic and hydrophilic contacts are coloured in red and blue, respectively. (b) Gel filtration analysis for interaction of JNK1 with MKP7-CD mutant F285D. Mutant F285D and JNK1 were eluted as monomers, with the molecular masses of ∼17 and 44 kDa, respectively. However, in contrast to the wild-type MKP7-CD, mutant F285D did not co-migrate with JNK1. (c) Pull-down assays of MKP7-CD by GST-tagged JNK1 mutants. The top panel shows the relative affinities of MKP7-CD to JNK1 mutants, with the affinity of wild-type JNK1 defined as 100%, the middle panel is the electrophoretic pattern of MKP7-CD and JNK1 mutants after GST pull-down assays. The protein amounts of MKP7-CD used are shown at the bottom. (d) Circular dichroism spectra for MKP7-CD wild type and mutants. Measurements were averaged for three scans. (e) Circular dichroism spectra for JNK1 wild type and mutants. Measurements were averaged for three scans. (f) Effects of mutations in MKP7-CD on the pNPP hydrolysis reaction (mean±s.e.m., n=3).

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