Development of an exchange assay for cytosolic glucocorticoid receptors using the synergistic effects of molybdate plus dithiothreitol

Journal Article · Thu Sep 01 00:00:00 EDT 1983 · Endocrinology; (United States)

OSTI ID:5745289

Kalimi, M; Hubbard, J R

A glucocorticoid receptor exchange assay has been developed for the accurate quantification of both free and steroid-bound receptors in rat liver cytosol. Hepatic cytosol from adrenalectomized rats was saturated in vitro with unlabeled corticosterone. Cytosol was subsequently treated with (/sup 3/H)dexamethasone (with and without 1000-fold cold dexamethasone) for 2-28 h at 4 C in the presence of 10 mM molybdate plus 5 mM dithiothreitol (DTT). Complete exchange occurred between 16-28 h in the presence of molybdate plus DTT. In control and 10 mM molybdate (alone) treated samples only about 50% exchange was achieved. In the presence of 5 mM DTT (alone) approximately 60-70% exchange was observed. The exchange assay (utilizing molybdate plus DTT) was also applied to hepatic cytosol of adrenalectomized rats injected with corticosterone in vivo and to samples prebound with unlabeled dexamethasone.

• Research Organization: Department of Physiology and Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond
• OSTI ID: 5745289
• Journal Information: Endocrinology; (United States), Vol. 113:3
• Country of Publication: United States
• Language: English

Related Subjects

62 RADIOLOGY AND NUCLEAR MEDICINE
DEXAMETHASONE
SYNERGISM
GLUCOCORTICOIDS
RECEPTORS
RATS
ADRENALECTOMY
BIOASSAY
IN VIVO
LIVER
MOLYBDATES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ADRENAL HORMONES
ANIMALS
BODY
CORTICOSTEROIDS
DIGESTIVE SYSTEM
GLANDS
HORMONES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
KETONES
LABELLED COMPOUNDS
MAMMALS
MEDICINE
MOLYBDENUM COMPOUNDS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PREGNANES
RODENTS
STEROID HORMONES
STEROIDS
SURGERY
TRANSITION ELEMENT COMPOUNDS
VERTEBRATES
550601* - Medicine- Unsealed Radionuclides in Diagnostics