Purification and crystallization of the human RXRalpha ligand-binding domain-9-cisRA complex.
The purification and crystallization of the stoichiometric complex of human RXRalpha ligand-binding domain (hRXRalpha LBD) bound to its natural ligand 9-cis retinoic acid (9-cisRA) are described. A three-step purification yields a pure and homogeneous complex. Based on the crystallization conditions of several other nuclear receptors, an exhaustive crystallization screening using carboxylic acids as precipitating agents was performed in association with the use of polyhydric alcohols acting as cosmotropic solutes. Crystals of the hRXRalpha LBD-9-cisRA complex grew in a tripartite mixture containing sodium formate, glycerol and propane-1,2-diol. Micro- and macroseeding were necessary to improve both the size and the quality of crystals in order to make them suitable for structure determination.
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Laboratoire de Biologie et Génomique Structurales, UPR 9004 CNRS, IGBMC CNRS/INSERM/ULP/Collège de France, Parc d'Innovation, 1 Rue Laurent Fries BP163, 67404 Illkirch CEDEX, France.
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D000077556:Alitretinoin; D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000302:isolation & purification; D047488:Retinoid X Receptors; D014157:Transcription Factors / Q000737:chemistry* / Q000302:isolation & purification; D014212:Tretinoin / Q000737:chemistry*
D018168:Receptors, Retinoic Acid; D047488:Retinoid X Receptors; D014157:Transcription Factors; D000077556:Alitretinoin; D014212:Tretinoin