FIG 5.

Autophosphorylation of BfmR in vitro by small phosphodonors. Purified BfmR protein (3 μM) was incubated with 10 mM acetylphosphate (A) or 10 mM carbamoyl phosphate (B) at 37°C for the indicated times, at which point reactions were stopped by the addition of SDS loading buffer. As a control, the last sample was heat shocked at 95°C for 5 min after 120 min incubation with the phosphodonor, prior to the addition of SDS loading buffer. Samples were then analyzed by Phos-tag gel electrophoresis and Western blotting.