Figure 2. Activity test of PknB_SA-KD.

PknB_SA-KD (25 ng) was incubated either with myelin basic protein (MBP; 1 µg) (A) or alone (B) together with γ³³-ATP, MnCl₂ and MgCl₂ for the time indicated. Position and size (kDa) of molecular weight markers are indicated on the left side. Phosphorylation of MBP (A) and autophosphorylation (B) are visualized by autoradiography using direct-exposure film. The phosphorylation rate is increasing as a function of time in both experiments, demonstrating that the purified PknB_SA-KD protein is active.