title: Recombinant human retinoic acid receptor alpha. Binding of DNA and synthetic retinoids to the protein expressed in Escherichia coli.
abstract: The human retinoic acid receptor alpha was expressed in Escherichia coli. The recombinant protein was found to be very unstable in several E. coli strains, probably due to proteolysis. Conditions were established to obtain reasonable amounts of active protein for ligand and DNA binding studies. The recombinant receptor showed the expected DNA binding activities in gel-retardation assays. Ligand binding properties were measured in a charcoal absorption assay. The dissociation constant for highly specific bound retinoic acid was found to be 0.67 nM. The affinity of several synthetic retinoids to the recombinant protein was determined and compared to their biological activity. Some of the values presented here differ significantly from those published earlier for the receptor or its isolated hormone-binding domain.
authors.affiliation: Department of Biochemistry, Medizinische Hochschule Hannover, Federal Republic of Germany.
authors.forename: S
authors.initials: S
authors.lastname: Keidel
authors.forename: E
authors.initials: E
authors.lastname: Rupp
authors.forename: M
authors.initials: M
authors.lastname: Szardenings
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002850:Chromatography, Gel; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D010957:Plasmids; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins / Q000378:metabolism; D012176:Retinoids / Q000138:chemical synthesis / Q000378:metabolism*; D014212:Tretinoin / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D008024:Ligands; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D012176:Retinoids; D014212:Tretinoin; D004247:DNA