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. 2013 Feb 28;25(2):649–661. doi: 10.1105/tpc.112.103184

Figure 5.

Figure 5.

Phosphorylation of CPK24 by CPK11 in Vitro.

(A) Coomassie blue–stained recombinant proteins of CPK11 and CPK24. Purified GST-CPK11 (left panel), GST-CPK24N (N-terminal, from Met-1 to Asn-195 residue; middle panel), GST-CPK24C (C-terminal, from Phe-196 to the end; middle panel), CPK24-His (right panel), GST-CPK11N (N-terminal, from Met-1 to Pro-153 residue; right panel), and GST-CPK11C (C-terminal, from Glu-154 to the end, right panel) are indicated with arrows in Coomassie blue–stained gels.

(B) Phosphorylation of the N terminus of CPK24 by CPK11. Each lane represents an independent reaction and contains 2 μg purified protein (kinase:substrate = 1:1). The arrowhead shows the autophosphorylation of CPK11, and the arrow shows the phosphorylation of CPK24 in the N terminus.

(C) CPK24 cannot phosphorylate CPK11 in vitro. The autoradiogram shows that neither of the truncated CPK11 variants (GST-CPK11N and GST-CPK11C) could be phosphorylated by CPK24 in vitro. The arrowhead shows the autophosphorylation of CPK24.