Figure 3: Chemoproteomics profiling of GSK3011724A.
From: Identification of KasA as the cellular target of an anti-tubercular scaffold
(a) A propylamine-tagged derivative of GSK3011724A (1, inset) was synthesized and covalently immobilized to NHS-activated sepharose. Beads were incubated with M. bovis BCG extract either in the presence of vehicle (DMSO) or GSK3011724A (10 μM, 40 μM). Proteins captured by the beads in both conditions were quantified by LC–MS/MS analysis. KasA, Pks10 and Pks11 were identified as potential targets of GSK3011724A by virtue of their reduced capturing in the presence of excess GSK3011724A. (b) Generation of IC50 values for KasA, Pks10 and Pks11. The chemoproteomic experiment was performed as in a but over a range of concentrations of the competing ‘free’ inhibitor GSK3011724A (2–0.003 μM for KasA, 40–0.16 μM for Pks10 and Pks11) and a structurally related inactive analogue, 2 (40–0.16 μM). Apparent dissociation constants for GSK3011724A were determined from two independent experiments.
