Fig. 4. Phosphorylation of BfmR in vitro induces dimerization.

(A) Phos-tag gel analysis of in vitro BfmR autophosphorylation. Purified BfmR protein was incubated with PA for one hour in Buffer S and combined with SDS loading buffer. In the last lane, the sample was heat inactivated at 95˚C for 10min before addition of SDS buffer. Gel was stained with SYPRO orange. (B) Analysis of BfmR dimerization via analytical SEC. Purified BfmR or BfmR^D58A was incubated with PA at 50μM (red) or no PA (blue) in Buffer S. Representative HPLC chromatograms are shown (representative of n=3).