Structure and function of cytosolic glucocorticoid receptors in rodent lymphoid cells: studies of receptor stability, subunit composition, and phosphorylation
The primary goal of these studies was to determine whether the cytosolic glucocorticoid receptor is dephosphorylated during the activation process in the intact cell. To address this question cytosolic complexes were purified from cells grown in medium containing (/sup 32/P)orthophosphoric acid or (/sup 32/S)methionine to allow quantitation of the number of phosphates and the amount of receptor protein, respectively. Steroid-binding proteins were identified by their specific association with the affinity label (/sup 3/H)dexamethasone 21-mesylate (DM). Attempts to purify nonactivated glucocorticoid-receptor complexes from rat thymocyte cytosol were unsuccessful due to the activity of an endogenous leupeptin-sensitive, calcium-activated protease which degraded the receptor to a form which was not recognized by the antibody. Nonactivated cytosolic complexes purified from WEHI-7 mouse thymoma cells using the BuGR1 monoclonal antibody contained a 90 kilodalton (90 kDa) non-steroid-binding subunit which could be separated from the 100 kDa steroid-binding subunit by SDS-PAGE. From these results the authors conclude that the nonactivated complex is a heteromeric structure which dissociates upon activation, and that activation of the complex in the intact cell does not result in a dephosphorylation of the 100 kDa steroid-binding protein.
- Research Organization:
- Dartmouth Coll., Hanover, NH (USA)
- OSTI ID:
- 5514424
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
LABELLED COMPOUNDS
BIOSYNTHESIS
RECEPTORS
BIOCHEMICAL REACTION KINETICS
PHOSPHORYLATION
ELECTROPHORESIS
GLUCOCORTICOIDS
LYMPHOCYTES
METHIONINE
MICE
MONOCLONAL ANTIBODIES
PHOSPHORIC ACID
PHOSPHORUS 32
PROTEINS
SULFUR 35
TRITIUM COMPOUNDS
ADRENAL HORMONES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
CORTICOSTEROIDS
DAYS LIVING RADIOISOTOPES
DRUGS
EVEN-ODD NUCLEI
HORMONES
HYDROGEN COMPOUNDS
HYDROXY COMPOUNDS
INORGANIC ACIDS
ISOTOPES
KETONES
KINETICS
LEUKOCYTES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MATERIALS
MEMBRANE PROTEINS
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS ISOTOPES
PREGNANES
RADIOISOTOPES
REACTION KINETICS
RODENTS
SOMATIC CELLS
STEROID HORMONES
STEROIDS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques