title: Purification of the human RARgamma ligand-binding domain and crystallization of its complex with all-trans retinoic acid.
abstract: A 28-kDa fragment (residues 178-423) of the human retinoic acid receptor gamma, hRARgamma D3E, encompassing the ligand-binding domain (LBD) was overproduced in Escherichia coli and purified as a monomer to more than 95% purity and homogeneity. The Kd for all-trans retinoic acid binding was 0.6 +/- 0.1 nM. Crystals of the LBD complexed with all-trans retinoic acid were grown at pH 7 from sodium acetate in the presence of detergents using the vapor diffusion method. They diffract to 2.0 A using a synchrotron radiation (lambda=0.91 A) and belong to the tetragonal space group P4(1)2(1)2 with unit cell parameters a=b=60.6 A and c=155.3 A, one monomer per asymmetric unit, a solvent content of ca. 33%, and a Vm value of approximately 2 A3/dalton.
authors.affiliation: Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP/Collège de France, Illkirch, France.
authors.forename: N
authors.initials: N
authors.lastname: Rochel
authors.forename: J P
authors.initials: JP
authors.lastname: Renaud
authors.forename: M
authors.initials: M
authors.lastname: Ruff
authors.forename: V
authors.initials: V
authors.lastname: Vivat
authors.forename: F
authors.initials: F
authors.lastname: Granger
authors.forename: D
authors.initials: D
authors.lastname: Bonnier
authors.forename: T
authors.initials: T
authors.lastname: Lerouge
authors.forename: P
authors.initials: P
authors.lastname: Chambon
authors.forename: H
authors.initials: H
authors.lastname: Gronemeyer
authors.forename: D
authors.initials: D
authors.lastname: Moras
mesh_terms: D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006801:Humans; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D016324:Sequence Tagged Sites; D014212:Tretinoin / Q000737:chemistry* / Q000378:metabolism*; D000097768:Retinoic Acid Receptor gamma
chemical_list: D017931:DNA Primers; D010446:Peptide Fragments; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D014212:Tretinoin; D000097768:Retinoic Acid Receptor gamma