Proteomics based approaches are emerging as useful tools to identify the targets of bioactive compounds and elucidate their molecular mechanisms of action. Here, we applied a chemical proteomic strategy to identify the peroxisome proliferator-activated receptor Î³ (PPARÎ³) as a molecular target of the pro-apoptotic agent 15-ketoatractyligenin methyl ester (compound 1). We demonstrated that compound 1 interacts with PPARÎ³, forms a covalent bond with the thiol group of C285 and occupies the sub-pocket between helix H3 and the Î²-sheet of the ligand-binding domain (LBD) of the receptor by Surface Plasmon Resonance (SPR), mass spectrometry-based studies and docking experiments. 1 displayed partial agonism of PPARÎ³ in cell-based transactivation assays and was found to inhibit the AKT pathway, as well as its downstream targets. Consistently, a selective PPARÎ³ antagonist (GW9662) greatly reduced the anti-proliferative and pro-apoptotic effects of 1, providing the molecular basis of its action. Collectively, we identified 1 as a novel PPARÎ³ partial agonist and elucidated its mode of action, paving the way for therapeutic strategies aimed at tailoring novel PPARÎ³ ligands with reduced undesired harmful side effects.
|Number of pages||14|
|Publication status||Published - 2017|