Autophosphorylation reactions of other hybrid sensor kinases, BarA and ArcB.

<p>(A) Phosphorylation sites in each domain of BarA and ArcB are shown. (B) Autophosphorylation reactions of wild-type BarA (WT) and the two mutants D718A and H861A were performed in the presence of 10 mM ATP, and the products were then analyzed by Phos-tag SDS-PAGE. The incubation times are shown above each lane. Each lane contained 2 μg of protein. Bands for each of the two site-specific phosphorylated forms were assigned and are shown by arrows on the right-hand side of the panel. (C) Autophosphorylation reactions of wild-type ArcB (WT) and the two mutants D576A and H717A were performed and analyzed in the same manner as BarA in B. (D) Values of the ratio of the phosphorylated forms to the total proteins in B and C were calculated by densitometry and are plotted versus the reaction times. Each plot was gained as average of three independent experiments using the same sample. Standard deviations were within almost 20%.