Fig. 6: The main target site of CPK1 at the N terminus of CNGC9 is Ser26. | Nature Communications

Fig. 6: The main target site of CPK1 at the N terminus of CNGC9 is Ser26.

From: CPK1 activates CNGCs through phosphorylation for Ca2+ signaling to promote root hair growth in Arabidopsis

Fig. 6

a, b LC-MS/MS spectrum show that the peptides LDSMDSR and KGFRKGSK contain the phospho-serines Ser26 (a) and Ser73 (b). c In vitro phosphorylation assay shows that the CPK1-mediated phosphorylation of CNGC9-NS26A was dramatically reduced, whereas the phosphorylation of CNGC9-NS73A was not obviously attenuated, compared to wild-type CNGC9-N. For the in vitro phosphorylation assays, the wild type and point-mutated CNGC9-N were cloned into the vector pCold-TF with a cleavable 6×His tag, and CPK1 was cloned into the vector pMAL-c5X with a MBP tag. The recombinants were expressed in E. coli and purified, and the recombinants with the tags retained were used for the in vitro protein phosphorylation assays. Three biological replicates were conducted for both the LC-MS/MS and the in vitro phosphorylation assays. Source data are provided as a Source Data file.

Back to article page