FIGURE 3.
In vitro phosphorylation of the GlgE orthologues from M. smegmatis and S. coelicolor. Recombinant M. smegmatis PknB kinase (PknBMs) and S. coelicolor AfsK kinase (AfsK) were expressed as His-tagged fusion proteins in E. coli, purified, and incubated with purified His-tagged M. smegmatis GlgE (GlgEMs) and S. coelicolor GlgE (GlgESc), respectively, in the presence of [γ-33P]ATP. GlgE and PknB proteins from M. tuberculosis were included as a positive control. Samples were separated by SDS-PAGE, stained with Coomassie Blue (upper panel), and visualized by autoradiography (lower panel). Marker indicates molecular mass markers.