Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Hermanson O, Glass C, Rosenfeld M (2002) Nuclear receptor coregulators: multiple modes of modification. Trends Endocrinol Metab 13:55–60
Nettles KW, Greene GL (2005) Ligand control of coregulator recruitment to nuclear receptors. Ann Review Physiol 67:309–333
Rosenfeld MG, Lunyak VV, Glass CK (2006) Sensors and signals: a coactivator/corepressor/epigenetic code for integrating signal-dependent programs of transcriptional response. Genes Dev 20:1405–1428
Pochetti G, Godio C, Mitro N et al (2007) Insights into the mechanism of partial agonism: crystal structures of the peroxisome proliferator-activated receptor gamma ligand-binding domainin the complex with two enatiomeric ligands. J Biol Chem 282:17314–17324
Pochetti G, Mitro N, Lavecchia A et al (2010) Structural insight into peroxisome proliferator-activated receptor gamma binding of two ureidofibrate-like enantiomers by molecular dynamics, cofactor interaction analysis, and site-directed mutagenesis. J Med Chem 53:4354–4366
Liu J, Knappenberger K, Kack H et al (2003) A homogeneous in vitro functional assay for estrogen receptors: coactivator recruitment. Mol Endocrinol 17:346–355
Darimont BD, Wagner R, Apriletti J et al (1998) Structure and specificity of nuclear receptor-coactivator interactions. Genes Dev 12:3343–3356
Acknowledgement
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Mitro, N., Godio, C., Crestani, M. (2013). Fluorescence Resonance Energy Transfer Techniques to Study Ligand-Mediated Interactions of PPARs with Coregulators. In: Badr, M., Youssef, J. (eds) Peroxisome Proliferator-Activated Receptors (PPARs). Methods in Molecular Biology, vol 952. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-155-4_16
Download citation
DOI: https://doi.org/10.1007/978-1-62703-155-4_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-154-7
Online ISBN: 978-1-62703-155-4
eBook Packages: Springer Protocols