Purification and crystallization of the heterodimeric complex of RARbeta and RXRalpha ligand-binding domains in the active conformation.
The ligand-binding domains of the retinoid X receptor alpha (RXRalpha) and of the retinoic acid receptor beta (RARbeta) were overexpressed separately and copurified in the heterodimeric form. Using a crystallization solution containing sodium formate and PEG 3350 as precipitant, the heterodimer was cocrystallized with the promiscuous ligand 9-cis-retinoic acid (9C-RA) and a 13-residue fragment of the nuclear receptor interaction domain (NID) of the transcriptional coactivator TRAP220. The crystals grew in the trigonal space group P3(1)21, with unit-cell parameters a = b = 115.7, c = 247.2 angstroms and two heterodimers per asymmetric unit. X-ray diffraction data were collected to 2.9 angstroms resolution. The structure was solved by molecular replacement and is currently being refined.
Pogenberg
Vivian
V
Centre de Biochimie Structurale, CNRS UMR 5048-Inserm U554, Faculté de Pharmacie, 15 Avenue Charles Flahault, 34093 Montpellier, France.
Guichou
Jean François
JF
Bourguet
William
W
D000077556:Alitretinoin; D000818:Animals; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism; D005561:Formates / Q000737:chemistry; D005822:Genetic Vectors; D006639:Histidine / Q000737:chemistry; D008024:Ligands; D051379:Mice; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D018168:Receptors, Retinoic Acid / Q000737:chemistry*; D000072482:Retinoic Acid Receptor alpha; D014212:Tretinoin / Q000737:chemistry; D014961:X-Ray Diffraction
D005561:Formates; D008024:Ligands; C000607072:Rara protein, mouse; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; C082925:retinoic acid receptor beta; C030544:formic acid; D000077556:Alitretinoin; D006639:Histidine; D014212:Tretinoin