Solubilization and biochemical characterization of the human myometrial corticotrophin-releasing hormone receptor
UNSPECIFIED. (1998) Solubilization and biochemical characterization of the human myometrial corticotrophin-releasing hormone receptor. MOLECULAR AND CELLULAR ENDOCRINOLOGY, 138 (1-2). pp. 185-198. ISSN 0303-7207Full text not available from this repository.
We have solubilized an active form of the myometrial corticotrophin-releasing hormone (CRH) receptor using 1% w/v digitonin. The solubilized receptor retains its capacity for high-affinity binding as demonstrated by Scatchard analysis, although there was a shift in dissociation constant (K-d) from 83.6 +/- 15-195 +/- 35 pM for the membrane-bound and soluble receptor respectively. There was no difference in the maximum binding site concentrations (B-max) of 13 +/- 5 and 21.5 +/- 6 fmol/mg protein for the membrane-bound and soluble receptor respectively. Sauvagine unlike CRH had no effect on radiolabeled CRH binding which suggests that the CRH-R2 receptor is not. present in the myometrium. The solubilized receptor did not retain guanine-nucleotide sensitivity. The isoelectric focusing (IEF) profile of the human myometrial CRH receptors was significantly different from that of the rat cerebral cortex. Furthermore, solubilization of human myometrial membrane proteins followed by gel filtration and SDS-PAGE revealed a specifically labeled protein with an apparent molecular weight of 42000-47000 kDa. Our results suggest that during solubilization the human myometrial CRH receptor is dissociated from the guanine nucleotide-binding protein (G(s)) and that high affinity binding for soluble CRH receptors is not dependent on the coupling of a guanine nucleotide-binding protein. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.
|Item Type:||Journal Article|
|Subjects:||Q Science > QH Natural history > QH301 Biology
R Medicine > RC Internal medicine
|Journal or Publication Title:||MOLECULAR AND CELLULAR ENDOCRINOLOGY|
|Publisher:||ELSEVIER SCI IRELAND LTD|
|Official Date:||16 March 1998|
|Number of Pages:||14|
|Page Range:||pp. 185-198|
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