DNA recognition by retinoic acid nuclear receptors.
Biophysics
Crystallization
DNA recognition
ESI MS
ITC
Protein expression and production
RAR-RXR-DNA complex
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2020
2020
Historique:
entrez:
4
5
2020
pubmed:
4
5
2020
medline:
24
6
2021
Statut:
ppublish
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.
Identifiants
pubmed: 32359647
pii: S0076-6879(20)30102-6
doi: 10.1016/bs.mie.2020.03.001
pii:
doi:
Substances chimiques
DNA-Binding Proteins
0
Receptors, Retinoic Acid
0
Retinoid X Receptors
0
Tretinoin
5688UTC01R
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
235-260Informations de copyright
© 2020 Elsevier Inc. All rights reserved.