title: RARbeta ligand-binding domain bound to an SRC-1 co-activator peptide: purification, crystallization and preliminary X-ray diffraction analysis.
abstract: Retinoids have demonstrated therapeutic efficacy in the treatment of acute promyelocytic leukaemia and in the chemoprevention of a large number of cancers. As the cellular signalling pathway of retinoids can be transduced by the three retinoic acid receptor (RAR) isotypes alpha, beta and gamma, the side effects of these treatments induced efforts to generate isotype-selective ligands. Despite knowledge of the crystal structures of RARalpha and RARgamma ligand-binding domains (LBDs), the rational design of such ligands has been hampered by the absence of RARbeta LBD structural data. Here, a strategy used to express a large-scale soluble fraction of the human RARbeta LBD suitable for biophysical analysis is reported, as well as a procedure for crystallizing it bound to a synthetic retinoid (TTNPB) with or without a co-activator peptide (SRC-1). Preliminary X-ray analysis revealed that both complexes crystallized in the orthorhombic space group P2(1)2(1)2(1). The unit-cell parameters are a = 47.81, b = 58.52, c = 92.83 A for the TTNPB-hRARbeta LBD crystal and a = 58.14, b = 84.07, c = 102.37 A when the SRC-1 peptide is also bound.
authors.affiliation: Department of Structural Biology and Genomics, Institut de Génétique et de Biologie Moléculaire et Cellulaire, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Kammerer
authors.forename: Pierre
authors.initials: P
authors.lastname: Germain
authors.forename: Ralf
authors.initials: R
authors.lastname: Flaig
authors.forename: Carole
authors.initials: C
authors.lastname: Peluso-Iltis
authors.forename: André
authors.initials: A
authors.lastname: Mitschler
authors.forename: Natacha
authors.initials: N
authors.lastname: Rochel
authors.forename: Hinrich
authors.initials: H
authors.lastname: Gronemeyer
authors.forename: Dino
authors.initials: D
authors.lastname: Moras
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D051548:Histone Acetyltransferases; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D056920:Nuclear Receptor Coactivator 1; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012176:Retinoids / Q000494:pharmacology; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D008024:Ligands; D010446:Peptide Fragments; D018168:Receptors, Retinoic Acid; D012176:Retinoids; D014157:Transcription Factors; C082925:retinoic acid receptor beta; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1