Purification and crystallization of the heterodimeric complex of RAR and RXRα ligand-binding domains in the active conformation
The ligand-binding domains of the retinoid X receptor α (RXRα) and of the retinoic acid receptor (RAR) 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 P3121, with unit-cell parameters a = b = 115.7, c = 247.2 Å and two heterodimers per asymmetric unit. X-ray diffraction data were collected to 2.9 Å resolution. The structure was solved by molecular replacement and is currently being refined.