SpaK/SpaR Two-component System Characterized by a Structure-driven Domain-fusion Method and in Vitro Phosphorylation Studies
Figure 4
In vitro phosphorylation studies of SpaK and SpaR.
A, B: SDS-PAGE of 6xHis-SpaK and 6xHis-SpaR in isolation or in combination and at various mass ratios, in the presence of ATP. A: Coomassie blue staining. B: Autoradiography; lane a: molecular weight markers. C: Phosphorimage analysis of SpaK incubated with [g-32P]-ATP (lane 1) followed by addition of 4 mM (lane 2), 10 mM (lane3), or 50 mM non-labeled (cold) ATP. D: PEI cellulose thin-layer chromatography of 6xHis-SpaK in isolation, or in combination with 6xHis-SpaR with and without EDTA.