TY - JOUR
T1 - Subunit composition of the molybdate-stabilized non-activated glucocorticoid receptor from rat liver
AU - Denis, M.
AU - Wikström, A. C.
AU - Gustafsson, J. A.
N1 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1988
Y1 - 1988
N2 - A monoclonal IgG 2a antibody directed against the activated rat liver glucocorticoid receptor (GR) was used to prepare an immunoaffinity matrix of high capacity. The molybdate-stabilized GR from rat liver cytosol was immunoadsorbed on this gel. A non-hormone-binding protein of Mr ≈ 90,000, as determined after denaturing gel electrophoresis, was eluted from this matrix following removal of molybdate and exposure to heat (25°C) and salt (0.15 M NaCl). Subsequently, the Mr ≈ 90,000 protein was purified to homogeneity using high-performance ion-exchange chromatography, covalently radiolabelled, and analyzed by high-performance size-exclusion chromatography and sucrose gradient ultracentrifugation. Hydrodynamic characterization indicates that, under our experimental conditions, the molybdate-stabilized rat liver GR (Rs ≈ 7.4nm, s20,w ≈9.1 S, calculated mol. wt Mr ≈ 285,000) includes one steroid-binding unit (Rs ≈ 5.5 nm, s20,w ≈ 3 S, calculated Mr ≈ 100,000) and a dimer of Mr ≈ 90,000 non-hormone-binding protein (Rs ≈ 6.9nm, s20,w ≈ 6.1 S, calculated native Mr ≈ 180,000).
AB - A monoclonal IgG 2a antibody directed against the activated rat liver glucocorticoid receptor (GR) was used to prepare an immunoaffinity matrix of high capacity. The molybdate-stabilized GR from rat liver cytosol was immunoadsorbed on this gel. A non-hormone-binding protein of Mr ≈ 90,000, as determined after denaturing gel electrophoresis, was eluted from this matrix following removal of molybdate and exposure to heat (25°C) and salt (0.15 M NaCl). Subsequently, the Mr ≈ 90,000 protein was purified to homogeneity using high-performance ion-exchange chromatography, covalently radiolabelled, and analyzed by high-performance size-exclusion chromatography and sucrose gradient ultracentrifugation. Hydrodynamic characterization indicates that, under our experimental conditions, the molybdate-stabilized rat liver GR (Rs ≈ 7.4nm, s20,w ≈9.1 S, calculated mol. wt Mr ≈ 285,000) includes one steroid-binding unit (Rs ≈ 5.5 nm, s20,w ≈ 3 S, calculated Mr ≈ 100,000) and a dimer of Mr ≈ 90,000 non-hormone-binding protein (Rs ≈ 6.9nm, s20,w ≈ 6.1 S, calculated native Mr ≈ 180,000).
UR - http://www.scopus.com/inward/record.url?scp=0023726212&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023726212&partnerID=8YFLogxK
U2 - 10.1016/0022-4731(88)90105-7
DO - 10.1016/0022-4731(88)90105-7
M3 - Article
C2 - 3386252
AN - SCOPUS:0023726212
SN - 0022-4731
VL - 30
SP - 271
EP - 276
JO - Journal of Steroid Biochemistry
JF - Journal of Steroid Biochemistry
IS - 1-6
ER -