title: A compound-based proteomic approach discloses 15-ketoatractyligenin methyl ester as a new PPARγ partial agonist with anti-proliferative ability.
abstract: 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 γ (PPARγ) as a molecular target of the pro-apoptotic agent 15-ketoatractyligenin methyl ester (compound 1). We demonstrated that compound 1 interacts with PPARγ, forms a covalent bond with the thiol group of C285 and occupies the sub-pocket between helix H3 and the β-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 PPARγ in cell-based transactivation assays and was found to inhibit the AKT pathway, as well as its downstream targets. Consistently, a selective PPARγ 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 PPARγ partial agonist and elucidated its mode of action, paving the way for therapeutic strategies aimed at tailoring novel PPARγ ligands with reduced undesired harmful side effects.
authors.affiliation: Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084, Fisciano, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Vasaturo
authors.affiliation: Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084, Fisciano, Italy.|PhD Program in Drug Discovery and Development, University of Salerno, Via Giovanni Paolo II 132, I-84084 Fisciano SA, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Fiengo
authors.affiliation: Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084, Fisciano, Italy.
authors.forename: Nunziatina
authors.initials: N
authors.lastname: De Tommasi
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Via Port'Arsa, 11, 82100 Benevento, Italy.
authors.forename: Lina
authors.initials: L
authors.lastname: Sabatino
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Via Port'Arsa, 11, 82100 Benevento, Italy.
authors.forename: Pamela
authors.initials: P
authors.lastname: Ziccardi
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Via Port'Arsa, 11, 82100 Benevento, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Colantuoni
authors.affiliation: Department of Organic Chemistry, University of Palermo, Viale delle Scienze - Parco d'Orleans II, 90128, Palermo, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Bruno
authors.affiliation: Department of Pharmacy, "Drug Discovery" Laboratory, University of Napoli "Federico II", Via D. Montesano, 49, 80131 Napoli, Italy.
authors.forename: Carmen
authors.initials: C
authors.lastname: Cerchia
authors.affiliation: Department of Pharmacy, "Drug Discovery" Laboratory, University of Napoli "Federico II", Via D. Montesano, 49, 80131 Napoli, Italy.
authors.forename: Ettore
authors.initials: E
authors.lastname: Novellino
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Via Port'Arsa, 11, 82100 Benevento, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Lupo
authors.affiliation: Department of Pharmacy, "Drug Discovery" Laboratory, University of Napoli "Federico II", Via D. Montesano, 49, 80131 Napoli, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lavecchia
authors.affiliation: Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084, Fisciano, Italy.|School of Medicine and Surgery, University of Salerno, via Salvatore Allende, 18, Baronissi, SA, Italy.
authors.forename: Fabrizio Dal
authors.initials: FD
authors.lastname: Piaz
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001665:Binding Sites; D049109:Cell Proliferation / Q000187:drug effects; D045786:Diterpenes, Kaurane / Q000494:pharmacology*; D004952:Esters / Q000737:chemistry / Q000494:pharmacology*; D057809:HEK293 Cells; D019073:HT29 Cells; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D008024:Ligands; D062105:Molecular Docking Simulation; D047495:PPAR gamma / Q000819:agonists*; D055550:Protein Stability; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D000077154:Rosiglitazone; D020349:Surface Plasmon Resonance; D013816:Thermodynamics; D045162:Thiazolidinediones / Q000494:pharmacology; D013997:Time Factors; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D045786:Diterpenes, Kaurane; D004952:Esters; D008024:Ligands; D047495:PPAR gamma; D045162:Thiazolidinediones; C587222:ent-kaurane 15-ketoatractyligenin methyl ester; D000077154:Rosiglitazone