Stimulation of specific binding of [H-3]-progesterone to bovine luteal cell-surface membranes: specificity of digitonin

Menzies, G.S. and Howland, Kevin and Rae, Michael T. and Bramley, Tony A. (1999) Stimulation of specific binding of [H-3]-progesterone to bovine luteal cell-surface membranes: specificity of digitonin. Molecular and Cellular Endocrinology, 153 (1-2). pp. 57-69. ISSN 0303-7207. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1016/S0303-7207(99)00091-X

Abstract

Non-genomic actions of progesterone have been described in the ovary, and luteal membranes of several species have been shown to possess specific binding sites for [H-3]-progesterone. However, binding of radiolabelled progesterone to luteal membranes was demonstrable only in the presence of digitonin. Digitonin is a non-ionic detergent which is thought to act by forming one-to-one complexes with certain sterols. It is also a cardiotonic agent, inhibiting (Na+-K+) ATPase activity by interaction with the extracellular (ouabain/K+) binding site. We therefore investigated which properties of digitonin were responsible for its stimulatory actions on progesterone binding to bovine luteal membranes. A range of compounds with detergent, cardiotonic and/or cholesterol-complexing activities were tested for their effects on [H-3]-progesterone binding to bovine luteal membrane fractions, and on haemolysis of rat erythrocytes. Stimulation of progesterone binding to luteal membranes was highly specific for digitonin, and a number of ionic and non-ionic detergents, cardenolides, saponins and cholesterol-complexing reagents tested failed either to stimulate [H-3]-progesterone binding to bovine luteal membranes in the absence of digitonin, or to inhibit binding specifically in the presence of digitonin. When digitonin was first reacted with excess cholesterol or pregnenolone to form the respective digitonides, stimulatory activity was greatly reduced, suggesting that the ability of digitonin to interact with (an) endogenous steroid(s) may be important in its action. High performance liquid chromatography (HPLC)-mass spectrometry of commercially available digitonin preparations indicated the presence of numerous minor impurities in most commercial digitonin preparations. Three major UV-absorbing peaks were isolated and characterised by mass spectrometry: all stimulated progesterone binding to bovine luteal membrane receptors in a dose-dependent manner, though to differing extents. Our data suggest that the unique action of digitonin on luteal membrane progesterone receptors is not related to its detergent or cardiotonic properties, but appears to be related to its ability to complex with membrane sterols. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.

Item Type: Article
Uncontrolled keywords: cow; corpus luteum; steroid; receptor; surface membrane
Subjects: Q Science
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: I.T. Ekpo
Date Deposited: 05 Apr 2009 22:26
Last Modified: 18 Jun 2014 08:59
Resource URI: http://kar.kent.ac.uk/id/eprint/16933 (The current URI for this page, for reference purposes)
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