Structural Insight into Peroxisome Proliferator-Activated Receptor γ Binding of Two Ureidofibrate-Like Enantiomers by Molecular Dynamics, Cofactor Interaction Analysis, and Site-Directed Mutagene

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• 作者：Giorgio Pochetti ; Nico Mitro ; Antonio Lavecchia ; Federica Gilardi ; Neva Besker ; Elena Scotti ; Massimiliano Aschi ; Nazzareno Re ; Giuseppe Fracchiolla ; Antonio Laghezza ; Paolo Tortorella ; Roberta Montan
• 刊名：Journal of Medicinal Chemistry
• 出版年：2010
• 出版时间：June 10, 2010
• 年：2010
• 卷：53
• 期：11
• 页码：4354-4366
• 全文大小：1124K
• 年卷期：v.53,no.11(June 10, 2010)
• ISSN：1520-4804

文摘

Molecular dynamics simulations were performed on two ureidofibrate-like enantiomers to gain insight into their different potency and efficacy against PPARγ. 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 PPARγ regulated promoters was confirmed by chromatin immunoprecipitation assays. Cell-based transcription assays show that PPARγ coactivator 1α (PGC-1α) and cAMP response element binding protein-binding protein (CBP) enhance the basal and ligand-stimulated receptor activity acting as coactivators of PPARγ, whereas the receptor interacting protein 140 (RIP140) and the nuclear corepressor 1 (N-CoR1) repress the transcriptional activity of PPARγ. 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.