Abstract
Previous gel filtration binding assay studies indicated that rat vascular smooth muscle cells contained corticoid receptor I and corticoid receptor II sites which could be distinguished on the basis of their relative affinities for aldosterone and dexamethasone. Ion-exchange chromatography experiments were designed to separate the two sites for further studies on their physical characteristics and role in vascular smooth muscle cell physiology. Cultured aortic cells were incubated with 5-10nM 3H steroid alone or in the presence of 10-fold non-radioactive steroid competitor for 30 min at 37°C. Following cell lysis, total cellular protein-bound steroid was isolated using Sephadex G-25 and applied to a DEAE-cellulose ion-exchange column. Three peaks of radioactivity were eluted using a 1-200 mM sodium phosphate gradient: peak T (30-38 mM), peak II (52-64 mM), and peak III (92-102 mM). Peaks I and II contained 60% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor II sites (dexfimethasone aldosterone). Peak III contained 40% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor I sites (aldosterone dexamethasone). These studies support the binding assay data on steroid specificity and relative proportion of type I and II sites. They also document the existence of type I and II corticoid receptors with different physicochemical characteristics in rat aortic smooth muscle cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 577-582 |
| Number of pages | 6 |
| Journal | Journal of Steroid Biochemistry |
| Volume | 22 |
| Issue number | 5 |
| DOIs | |
| State | Published - 1985 |
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ASJC Scopus subject areas
- Biochemistry
- Endocrinology
Cite this
Physical separation of aortic corticoid receptors with type I and type II specificities. / Nichols, Nancy R.; Nguyen, Hanh H.; Meyer, Walter.
In: Journal of Steroid Biochemistry, Vol. 22, No. 5, 1985, p. 577-582.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Physical separation of aortic corticoid receptors with type I and type II specificities
AU - Nichols, Nancy R.
AU - Nguyen, Hanh H.
AU - Meyer, Walter
PY - 1985
Y1 - 1985
N2 - Previous gel filtration binding assay studies indicated that rat vascular smooth muscle cells contained corticoid receptor I and corticoid receptor II sites which could be distinguished on the basis of their relative affinities for aldosterone and dexamethasone. Ion-exchange chromatography experiments were designed to separate the two sites for further studies on their physical characteristics and role in vascular smooth muscle cell physiology. Cultured aortic cells were incubated with 5-10nM 3H steroid alone or in the presence of 10-fold non-radioactive steroid competitor for 30 min at 37°C. Following cell lysis, total cellular protein-bound steroid was isolated using Sephadex G-25 and applied to a DEAE-cellulose ion-exchange column. Three peaks of radioactivity were eluted using a 1-200 mM sodium phosphate gradient: peak T (30-38 mM), peak II (52-64 mM), and peak III (92-102 mM). Peaks I and II contained 60% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor II sites (dexfimethasone aldosterone). Peak III contained 40% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor I sites (aldosterone dexamethasone). These studies support the binding assay data on steroid specificity and relative proportion of type I and II sites. They also document the existence of type I and II corticoid receptors with different physicochemical characteristics in rat aortic smooth muscle cells.
AB - Previous gel filtration binding assay studies indicated that rat vascular smooth muscle cells contained corticoid receptor I and corticoid receptor II sites which could be distinguished on the basis of their relative affinities for aldosterone and dexamethasone. Ion-exchange chromatography experiments were designed to separate the two sites for further studies on their physical characteristics and role in vascular smooth muscle cell physiology. Cultured aortic cells were incubated with 5-10nM 3H steroid alone or in the presence of 10-fold non-radioactive steroid competitor for 30 min at 37°C. Following cell lysis, total cellular protein-bound steroid was isolated using Sephadex G-25 and applied to a DEAE-cellulose ion-exchange column. Three peaks of radioactivity were eluted using a 1-200 mM sodium phosphate gradient: peak T (30-38 mM), peak II (52-64 mM), and peak III (92-102 mM). Peaks I and II contained 60% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor II sites (dexfimethasone aldosterone). Peak III contained 40% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor I sites (aldosterone dexamethasone). These studies support the binding assay data on steroid specificity and relative proportion of type I and II sites. They also document the existence of type I and II corticoid receptors with different physicochemical characteristics in rat aortic smooth muscle cells.
UR - http://www.scopus.com/inward/record.url?scp=0021870412&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021870412&partnerID=8YFLogxK
U2 - 10.1016/0022-4731(85)90208-0
DO - 10.1016/0022-4731(85)90208-0
M3 - Article
C2 - 2989617
AN - SCOPUS:0021870412
VL - 22
SP - 577
EP - 582
JO - Journal of Steroid Biochemistry and Molecular Biology
JF - Journal of Steroid Biochemistry and Molecular Biology
SN - 0960-0760
IS - 5
ER -