Extended Data Fig. 2: PPK2 phosphorylates apo- and holo-phyA in vitro. | Nature Plants

Extended Data Fig. 2: PPK2 phosphorylates apo- and holo-phyA in vitro.

From: Liquid–liquid phase separation of TZP promotes PPK-mediated phosphorylation of the phytochrome A photoreceptor

Extended Data Fig. 2

In vitro kinase assays showing that PPK2-catalyzed phosphorylation of holo- and apo-phyA with different concentrations of phyA (a,c) or for different times (b,d). For each panel, CBB staining (top), and autoradiographs (bottom) are shown, with zinc fluorescence (Zinc) shown in the middle of a and b. PhyA-GST proteins were expressed in and purified from Saccharomyces cerevisiae BJ2168, and phycocyanobilin (PCB) extracted from Spirulina was added to the supernatant at a final concentration of 10 µM in the dark to allow the assembly of holo-phyA proteins in vitro. The holo-phyA proteins were exposed to 15 min of FR light alone, or 15 min of FR light immediately followed by 15 min of R light to allow holo-phyA to form Pr or Pfr forms in vitro, respectively.

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