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Identification of Ser-39 and Ser-40 in hCKβ as the major sites of PKA phosphorylation.

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posted on 05.05.2016, 07:52 by Ching Ching Chang, Ling Ling Few, Manfred Konrad, Wei Cun See Too

A) Schematic representation of potential sites of PKA phosphorylation in hCKβ as determined by mass spectrometry. In vitro phosphorylation of B) GST-hCKβ and GST-∆42NhCKβ recombinant fusion proteins, C) hCKβ, S39AhCKβ, S40AhCKβ, S42AhCKβ, S39A/S42AhCKβ, S40A/S42AhCKβ, and D) S39A/S40AhCKβ mutant proteins. The pictures are representative of at least two independent experiments. All CKβ mutant proteins were in vitro phosphorylated with 80 U PKA as described in the experimental procedures. Five micrograms of phosphorylated CKβ were loaded in each lane, and phosphorylation was detected with Pro-Q Diamond phosphoprotein stain. Total protein was stained with either Coomassie blue (B & D) or SYPRO® Ruby stain (C).