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Methods Mol Biol. 2013;952:219-27. doi: 10.1007/978-1-62703-155-4_16.

Fluorescence resonance energy transfer techniques to study ligand-mediated interactions of PPARs with coregulators.

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Laboratorio Giovanni Galli di Biochimica e Biologia Molecolare del Metabolismo-Spettrometria di Massa, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milan, Italy.


The capacity to induce the association of peroxisome proliferator-activated receptors (PPARs) with different transcriptional coregulators is determined by the peculiar 3D-structure that the receptors adopt when bound with a specific ligand. The fluorescence resonance energy transfer assay is a technique widely used to evaluate coregulator recruitment to nuclear receptors induced by ligands. With this assay it is possible to quantitatively determine the interaction and the affinity of coregulators with PPARs when these receptors are complexed with ligands. Here, we describe the use of this technique to assess the preferential interaction and the affinity of PPAR╬│ with coregulators as a function of the chemical structure of the bound ligand.

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