FIG. 3.

Comparison of protein profile and in vivo phosphorylation patterns in M. pneumoniae wild-type and mutant cells. Whole-cell extracts of the M. pneumoniae wild type and different mutant strains were analyzed by SDS-PAGE and stained with Pro-Q Diamond (Invitrogen) and Flamingo fluorescent dye (Bio-Rad) for visualization of phosphoproteins and all proteins, respectively. Ten μg of extract was applied to each lane. Interesting protein bands were cut out and identified by MS. Protein bands with changes in the phosphorylation signal or the protein amount are indicated by arrows. Total protein (A) (Flamingo fluorescent stain); phosphoproteins (B) (Pro-Q Diamond stain). (C) Relative quantification of phosphosignal intensity. The graph shows the changes relative to the phosphorylation signal of the corresponding protein in the M. pneumoniae wild type. Error bars indicate the standard deviation (based on three independent experiments). Note that the protein amount of HMW1-3 and P1 is also decreased in the prkC mutant.