Fig. 7

Ligand binding mutation causes global hyperphosphorylation of specific and non-specific targets. a TMT intensities of phosphopetides in RvΔB - pristinamycin (PknB depleted sample, labeled with TMT 127) or in RvΔB::B (complemented with 3F-PknB, labeled with TMT 128) or RvΔB::B-GM (complemented with 3F-PknB-GM, labeled with TMT131), with respect to RvΔB +pristinamycin as the reference comparator (control strain, labeled with TMT 126). The intensities of phosphopeptides in each case were normalized with respect to the corresponding absolute protein intensities and the values were converted to log2 values. Data were sorted with respect to RvΔB::B sample and heatmap of the data were generated using online tool Morpheus. b The 257 phosphopeptides detected in TMT experiment belonged to 147 unique proteins, which were classified according to their functional category with reference to mycobrowser database. The phosphopeptides were categorized as probable PknB substrates if the TMT log₂ phosphointensity upon depletion was <−0.32 and upon complementation was >1. 111 phosphopeptides were classified as probable PknB substrates, which belonged to 73 unique proteins. The 73 PknB targets were also functionally characterized according to mycobrowser database. c Normalized TMT intensities of all 111 phosphopeptides which are probable PknB targets were converted to log₂ values and data were sorted with respect to RvΔB::B. Heatmap was generated using online tool Morpheus. d Normalized TMT intensities of 5 phosphopeptides each belonging to cluster 1, cluster 2, and cluster 3. e Normalized TMT intensities of PknB-dependent tyrosine phosphorylations are represented. Source data are provided as a Source Data file