title: Structural insight into peroxisome proliferator-activated receptor gamma binding of two ureidofibrate-like enantiomers by molecular dynamics, cofactor interaction analysis, and site-directed mutagenesis.
abstract: Molecular dynamics simulations were performed on two ureidofibrate-like enantiomers to gain insight into their different potency and efficacy against PPARgamma. The partial agonism of the S enantiomer seems to be due to its capability to stabilize different regions of the receptor allowing the interaction with both coactivators and corepressors as shown by fluorescence resonance energy transfer (FRET) assays. The recruitment of the corepressor N-CoR1 by the S enantiomer on two different responsive elements of PPARgamma regulated promoters was confirmed by chromatin immunoprecipitation assays. Cell-based transcription assays show that PPARgamma coactivator 1alpha (PGC-1alpha) and cAMP response element binding protein-binding protein (CBP) enhance the basal and ligand-stimulated receptor activity acting as coactivators of PPARgamma, whereas the receptor interacting protein 140 (RIP140) and the nuclear corepressor 1 (N-CoR1) repress the transcriptional activity of PPARgamma. We also tested the importance of the residue Q286 on the transcriptional activity of the receptor by site-directed mutagenesis and confirmed its key role in the stabilization of helix 12. Molecular modeling studies were performed to provide a molecular explanation for the different behavior of the mutants.
authors.affiliation: Istituto di Cristallografia, Consiglio Nazionale delle Ricerche, Montelibretti, 00015 Monterotondo Stazione, Roma, Italia.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Pochetti
authors.forename: Nico
authors.initials: N
authors.lastname: Mitro
authors.forename: Antonio
authors.initials: A
authors.lastname: Lavecchia
authors.forename: Federica
authors.initials: F
authors.lastname: Gilardi
authors.forename: Neva
authors.initials: N
authors.lastname: Besker
authors.forename: Elena
authors.initials: E
authors.lastname: Scotti
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Aschi
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Re
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Fracchiolla
authors.forename: Antonio
authors.initials: A
authors.lastname: Laghezza
authors.forename: Paolo
authors.initials: P
authors.lastname: Tortorella
authors.forename: Roberta
authors.initials: R
authors.lastname: Montanari
authors.forename: Ettore
authors.initials: E
authors.lastname: Novellino
authors.forename: Fernando
authors.initials: F
authors.lastname: Mazza
authors.forename: Maurizio
authors.initials: M
authors.lastname: Crestani
authors.forename: Fulvio
authors.initials: F
authors.lastname: Loiodice
mesh_terms: D001583:Benzoxazoles / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D002087:Butyrates / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D056970:Co-Repressor Proteins / Q000378:metabolism; D006801:Humans; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D047495:PPAR gamma / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D000077154:Rosiglitazone; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D045162:Thiazolidinediones / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: D001583:Benzoxazoles; D002087:Butyrates; D056970:Co-Repressor Proteins; D047495:PPAR gamma; D045162:Thiazolidinediones; D000077154:Rosiglitazone