Conservation of the DNA binding domain and other properties between porcine and rat glucocorticoid receptors

• Published in: Journal of steroid biochemistry Vol. 24; no. 6; pp. 1097 - 1103
• Main Authors: Marks, Andrew R., Moore, David D., Buckley, Douglas I., Gametchu, Bahiru, Goodman, Howard M.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier B.V 01.06.1986; New York, NY Pergamon
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Antibodies, Monoclonal; Base Sequence; Binding and carrier proteins; Biological and medical sciences; Chromatography, Affinity; Collodion; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Immunologic Techniques; Male; Mutation; Proteins; Rats; Receptors, Cell Surface - genetics; Receptors, Cell Surface - isolation & purification; Receptors, Glucocorticoid - genetics; Receptors, Glucocorticoid - isolation & purification; Receptors, Somatotropin; Species Specificity; Swine; Rat; Steroid hormone; Rodentia; Chemical structure; Gene expression; Glucocorticoid; Pig; Binding protein; Vertebrata; Biological fixation; Mammalia; DNA; Hormone receptor complex; Receptor protein; Artiodactyla; Ungulata
• ISSN: 0022-4731
• DOI: 10.1016/0022-4731(86)90369-9

Activated glucocorticoid receptor protein (GCR) was partially purified from porcine liver cytosol by sequential chromatography on phosphocellulose and DNA-cellulose using a modification of a protocol developed for purification of rat GCR [1–2]. This partially purified preparation, when separated by SDS-polyacrylamide gel electrophoresis and immunoblotted, indicated that a Mr = 94,000 protein band cross-reacts with a monoclonal antibody against rat GCR [3]. A nitrocellulose filter binding assay showed that both the partially purified porcine and rat GCRs interact specifically with a cloned synthetic 24 base pair deoxyoligonucleotide containing the GCR binding sequence in the first intron of the human growth hormone (hGH) gene [4]. This specific protein-DNA interaction is blocked by a single base pair change in the binding site. All three putative domains of the GCR molecule: the steroid binding, immunoreactive, and DNA binding have been conserved between two divergent species.