Figure 4
From: Analysis of crystal structure of Arabidopsis MPK6 and generation of its mutants with higher activity
In vitro kinase activity assays of MPK6 and its mutants with or without the active MKK5 (MKK5DD).
The recombinant proteins for the active MKK5 and MPK6 mutants were expressed in E. coli and purified and subjected to an in-solution kinase assay using myelin basic protein as the substrate. MBP phosphorylation was shown in top panel. The coomassie bright blue (CBB)-stained gels showed that equal amounts of the MPK6 mutants (middle panel) and MBP (bottom panel) were used in each reaction. (A) Phosphorylation of the MBP substrate by the MPK6 mutants in the presence of active MKK5 (MKK5DD). (B) Phosphorylation of the MBP substrate by the MPK6 mutants in the absence of active MKK5 (MKK5DD).
