FIG. 5.
Autophosphorylation of truncated His6-CbbSRT189. The phosphorylation reaction was carried out in a final volume of 100 μl with 30 μg of protein (380 pmol) in the presence of 20 μM [γ-32P]ATP. At the indicated time points, 10 μl of the reaction mixture was withdrawn, quenched with 3× SDS loading dye-buffer, and stored on ice until electrophoretic separation. (A) Time course of autophosphorylation of wild-type His6-CbbSRT189. (B) Time course of autophosphorylation of D696N His6-CbbSRT189. (C) The concentration of phosphorylated product, obtained from a calibration curve as described in Materials and Methods, was plotted against the reaction time for wild-type His6-CbbSRT189 (▵) and D696N His6-CbbSRT189 (▪).