Mycobacterium tuberculosis VadK required for regulation of the methylcitrate cycle and virulence

We performed co-immunoprecipitation experiments using WT Mtb transformed with a plasmid encoding a FLAG-tagged VadK under a constitutive promoter (VadKMtb). We immunoprecipitated from whole cell lysates of WT and VadKMtb grown in standard 7H9 media with and without propionate to identify interactions that are important during growth in propionate. By applying a stringent cut-off to filter out nonspecific binding (proteins with peptide spectrum matches (PSMs) of each replicate ≤1 in the WT control and ≥50 in VadKMtb).