Fluorescence Resonance Energy Transfer Techniques to Study Ligand-Mediated Interactions of PPARs with Coregulators
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 PPARγ with coregulators as a function of the chemical structure of the bound ligand.
Key wordsNuclear receptors Peroxisome proliferator-activated receptors Coactivators Corepressors Protein–protein interaction Fluorescence resonance energy transfer
This work was supported by grants from the European Commission (LSHM-CT2006-037498), the Giovanni Armenise-Harvard Career Development Award and the Cariplo Foundation (2008.2511 and 2009.2727). We are grateful to Miss Elda Desiderio Pinto for invaluable administrative support.
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