Figure 3

CrMPK3 shows autophosphorylation as well as MBP phosphorylation. (A) CrMPK3 expressed in E. coli, BL21 strain as GST fusion was analyzed for autophosphorylation, as well as phosphorylation of MBP in an in-solution kinase assay using radiolabled ATP. GST alone was used in the assay and showed no auto- and substrate phosphorylation. (B). An in-gel kinase assay with decreasing concentrations of GST-CrMPK3 using MBP as substrate. The amount of recombinant proteins used was 1 μg; 0.1 μg; 0.01 μg; 0.001 μg from lane1 to 4. The phosphorylated MBP was visualized by autoradiography. The experiments have been repeated three times with similar result.