Abstract
The role of certain amino acids in the interactions of ligands with their cognate nuclear receptors is usually achieved by the resolution of the crystal structure of the receptor complexed with the ligand. As a complementary functional approach, site-directed mutagenesis, a technique broadly used in molecular biology, allows the assessment of the role of a specific amino acid in determining the interaction with a specific ligand. This method makes it possible to evaluate several mutations of a key amino acid for ligand binding and to determine the relationship between protein structure and ligand interaction. Here, we describe an application of this technique to evaluate different point mutations on the transcriptional activity of peroxisome proliferator-activated receptor γ (PPARγ) in the absence or presence of chemically different ligands.
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References
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Acknowledgments
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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Mitro, N., Gilardi, F., Giudici, M., Godio, C., Scotti, E., Crestani, M. (2013). Site-Directed Mutagenesis to Study the Role of Specific Amino Acids in the Ligand Binding Domain of PPARs. 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_9
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DOI: https://doi.org/10.1007/978-1-62703-155-4_9
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-154-7
Online ISBN: 978-1-62703-155-4
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