Monoclonal antibody to the rat glucocorticoid receptor. Relationship between the immunoreactive and DNA-binding domain

Journal Article · Wed Sep 25 04:00:00 EDT 1985 · J. Biol. Chem.; (United States)

OSTI ID:6045484

Eisen, L P; Reichman, M E; Thompson, E B; Gametchu, B; Harrison, R W; Eisen, H J

The region of the glucocorticoid receptor that reacted with a monoclonal antibody (BUGR-1) was identified. In order to identify the immunoreactive region, the rat liver glucocorticoid receptor was subjected to limited proteolysis; immunoreactive fragments were identified by Western blotting. The monoclonal antibody reacted with both the undigested Mr approximately 97,000 receptor subunit and a Mr approximately 45,000 fragment containing the steroid-binding and DNA-binding domains. Digestion by trypsin also produced two steroid-binding fragments of Mr approximately 27,000 and 31,000 which did not react with the antibody and an immunoreactive Mr approximately 16,000 fragment. This Mr approximately 16,000 fragment was shown to bind to DNA-cellulose, indicating that it contained a DNA-binding domain of the receptor. The undigested receptor must have steroid associated with it to undergo activation to a DNA-binding form. However, the Mr approximately 16,000 immunoreactive fragment binds to DNA-cellulose even if it is obtained by digestion of the steroid-free holoreceptor which does not itself bind to DNA.

• Research Organization: National Cancer Institute, Bethesda, MD
• OSTI ID: 6045484
• Journal Information: J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 21; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADRENAL HORMONES
ANIMALS
ANTIBODIES
BIOCHEMISTRY
CHEMISTRY
CORTICOSTEROIDS
DEXAMETHASONE
DNA
GLUCOCORTICOIDS
HORMONES
HYDROXY COMPOUNDS
KETONES
LABELLED COMPOUNDS
MAMMALS
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PREGNANES
RATS
RECEPTORS
RODENTS
STEROID HORMONES
STEROIDS
TRITIUM COMPOUNDS
VERTEBRATES