Figure 2. in vitro phosphorylation of VicK in the combined presence of Mn²⁺ and other various metal cations.

VicK (1 µM) was incubated in 100 mM Tris-HCl, pH 7.5 containing 1 mM MnCl₂ plus 1 mM of the designated cation and 0.10 µM [γ-³²P] ATP at room temperature for 15 minutes. The relative autophosphorylation of VicK was quantified using Image Quant 5.0 software (Molecular Dynamics). The sample containing only VicK and MnCl₂ was set at 100% for comparison and the results are shown in the histogram above the scanned gel. The gels shown are representative of at least three independent experiments. Error bars represent ± std. errors of the average phosphorylation values derived from at least 3 independent experiments.