Fiche publication
Date publication
janvier 2020
Journal
Methods in enzymology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr ROCHEL-GUIBERTEAU Natacha
,
Dr OSZ-PAPAI Judit
Tous les auteurs :
Peluso-Iltis C, Osz J, Rochel N
Lien Pubmed
Résumé
Retinoic acid receptors (RARs) heterodimerize with retinoid X receptors (RXRs) to regulate gene expression. The heterodimer recognizes the genome via a large and diverse repertoire of DNA response elements. Assessing the binding mode of RAR and RXR with various DNA response elements is important for understanding how they select their binding site and how DNA sequence and topology allosterically regulate RAR function. A number of complementary assays are often employed for analysis of the binding mode. To biochemically and structurally characterize RAR and RXR-DNA complexes, we describe how to express and purify RAR and RXR-DNA binding domains (DBDs) and multidomain constructs. We also describe the use of electrospray ionization mass spectrometry (ESI MS) and isothermal titration calorimetry (ITC) that give information about stoichiometry and binding affinity, as well as our approaches for co-crystallization of RAR and RXR DBDs with DNA.
Mots clés
Biophysics, Crystallization, DNA recognition, ESI MS, ITC, Protein expression and production, RAR-RXR-DNA complex
Référence
Meth. Enzymol.. 2020 ;637:235-260
