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. 2014 Jul 18;166(1):314–326. doi: 10.1104/pp.114.240226

Figure 5.

Figure 5.

In vitro phosphorylation of peptide arrays by CPK13. A, Representative autoradiography of KAT1, KAT2, and AKT2 peptide arrays after phosphorylation by recombinant CPK13. Peptide arrays contain 52 peptides (15–18 amino acid each) corresponding to the wild-type KAT1 sequence (including 11 in common with KAT2), 41 peptides corresponding to the wild-type KAT2 sequence (different from KAT1), and 60 peptides corresponding to the wild-type AKT2 sequence. The list of these peptides is given in Supplemental Table S1. A number of mutant peptides are spotted between some wild-type peptides (not listed in Supplemental Table S1). Black spots correspond to phosphorylated peptides. B, Schematic drawing of the secondary structure of the KAT2 Shaker channel subunit, highlighting the putative sites phosphorylated by CPK13 (white dots) as deduced from the on-chip assay of kinase activity shown in A. ext, cyt, and mb stand for extracellular, cytoplasmic side, and membrane; S1 to S6, P, CNBD, and KHA stand for transmembrane segments 1 to 6, pore domain, putative cyclic nucleotide-binding domain, and domain rich in hydrophobic and acidic residues, respectively.