TY - JOUR
T1 - Interaction of rat liver glucocorticoid receptor with heparin
AU - Weisz, Alessandro
AU - Puca, Giovanni Alfredo
AU - Masucci, Maria Teresa
AU - Masi, Carlo
AU - Pagnotta, Rita
AU - Petrillo, Antonella
AU - Sica, Vincenzo
PY - 1984
Y1 - 1984
N2 - When rat liver cytosol containing [3H]dexamethasone-glucocorticoid receptor complex is exposed to immobilized heparin (Sepharose-heparin; Seph-hep) the steroid receptor complex binds to the substituted Sepharose avidly [Kd = 3.5 (±1.7) × 10-10 M], and 80-90% of the receptor present is adsorbed to the solid phase after 40 min at 0°C. The binding is enhanced by Mn2+ (10 mM) and Mg2+, whereas Ca2+ and Sr2+ are ineffective. Sodium molybdate (10 mM) does not influence the reaction but enhances receptor stability. Moreover, binding of the receptor to Seph-hep is dependent on the ionic strength of the medium, because binding is totally reversed by 300 mM KCl. The bound [3H]dexamethasone-receptor complex can be recovered from Seph-hep with solutions (4 mg/mL) of heparin (95% release), dextran sulfate (88%), and chondroitin sulfate (63%); total calf liver RNA is less effective (9%), whereas dextran, D-glucosamine, N-acetyl-D-glucosamine, D-glucuronic acid, and sheared calf thymus DNA are totally ineffective (3H] dexamethasone-receptor bound to the resin.
AB - When rat liver cytosol containing [3H]dexamethasone-glucocorticoid receptor complex is exposed to immobilized heparin (Sepharose-heparin; Seph-hep) the steroid receptor complex binds to the substituted Sepharose avidly [Kd = 3.5 (±1.7) × 10-10 M], and 80-90% of the receptor present is adsorbed to the solid phase after 40 min at 0°C. The binding is enhanced by Mn2+ (10 mM) and Mg2+, whereas Ca2+ and Sr2+ are ineffective. Sodium molybdate (10 mM) does not influence the reaction but enhances receptor stability. Moreover, binding of the receptor to Seph-hep is dependent on the ionic strength of the medium, because binding is totally reversed by 300 mM KCl. The bound [3H]dexamethasone-receptor complex can be recovered from Seph-hep with solutions (4 mg/mL) of heparin (95% release), dextran sulfate (88%), and chondroitin sulfate (63%); total calf liver RNA is less effective (9%), whereas dextran, D-glucosamine, N-acetyl-D-glucosamine, D-glucuronic acid, and sheared calf thymus DNA are totally ineffective (3H] dexamethasone-receptor bound to the resin.
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M3 - Article
C2 - 6509026
AN - SCOPUS:0021733615
VL - 23
SP - 5393
EP - 5397
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 23
ER -