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. 1998 Nov 10;95(23):13976–13981. doi: 10.1073/pnas.95.23.13976

Figure 1.

Figure 1

Recombinant oat and algal phytochromes possess protein kinase activity. (A) Autoradiograph of Pr and Pfr forms of PCB adducts of recombinant oat phytochrome A (AsphyA-ST; 0.5 μg) after incubation with radiolabeled ATP under standard kinase conditions in the presence (+) or absence (−) of 2 μg histone H1 and/or 2 mM pyrophosphate (PPi) was obtained as described in Materials and Methods. The relative amount of phosphorylation indicated below the autoradiographs was obtained by using imagequant software (Molecular Dynamics). Dots on the left side of the gel represent molecular mass standards with 197 kDa, 117 kDa, 89 kDa, and 52 kDa (top to bottom). (B) An autoradiograph of algal phytochrome (Mcphy1b-ST; 1 μg) assayed identically as AsphyA-ST (above). (C) Hydrolytic stability of autophosphorylated phytochromes examined by treatment of poly(vinylidene difluoride) membranes with neutral (50 mM Tris⋅HCl, pH 7.5), basic (3 M KOH), or acidic (1 M HCl) solutions for 2.5 hr at room temperature with gentle shaking.