FIG. 5.

Western blot for the detection of the different phosphorylation forms of HPr. Crude extracts of M. pneumoniae grown in the presence of different carbon sources (1% final concentration) were separated by using native gels. For each condition tested, a parallel aliquot was incubated for 10 min at 70°C to hydrolyze the heat-labile HPr(His∼P). The different HPr species [HPr, HPr(His∼P), HPr(Ser-P), and HPr(His∼P)(Ser-P)] were detected by using polyclonal rabbit antibodies raised against M. pneumoniae His₆-tagged HPr. Ten micrograms of extract was applied to each lane.