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- W2854323797 abstract "We have subjected homogenates of mid-luteal phase equine CL to fractionation on continuous sucrose density gradients and have assayed gradient fractions for progesterone content, for specific binding of [3H]progesterone and [125I]-labeled LH, and for a number of marker activities for the major intracellular organelles. Progesterone content (a reflection of progesterone synthesis) equilibrated in membrane fractions with a buoyant density of 20–24% sucrose (1.08–1.10 g/cm3). In contrast, specific binding of [3H]-labeled progesterone equilibrated at a buoyant density of 25–34% sucrose (1.10–1.14 g/ cm3), similar to that of markers for luteal cell surface membranes (LH receptor, alkaline phosphatase) but distinct from that of other major luteal cell organelles. Binding of radiolabeled progesterone to dextran-charcoal-stripped equine luteal membrane fractions pooled from the 20–24% sucrose (1.08–1.10 g/cm3) region of the gradients was low, but it could be stimulated markedly and in a dose-dependent fashion by digitonin. Other detergents and cardiotonic steroids had no effect. In the presence of digitonin, [3H]progesterone binding increased with increasing membrane concentration, and binding was dependent on the pH, duration, and temperature of incubation. Unlabeled progesterone competed for the binding of [3H]progesterone with a concentration required to reduce specific binding by 50% (IC50) of 70 nM, whereas 17-hydroxyprogesterone, dihydrotestosterone, and testosterone had lower potencies (IC50, 900–3130 nM). Dehydroepiandrosterone (14 μM), pregnenolone, and androstenedione were even less effective (250 μM); and millimolar concentrations of estrogen, steroid conjugates, corticosteroids, cholesterol, inhibitors of luteal steroidogenic enzymes, and the progesterone receptor antagonist RU486 failed to compete. These results indicate the presence of specific, high-affinity binding sites for progesterone associated with luteal cell-surface membranes of the mare." @default.
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- W2854323797 date "1995-01-01" @default.
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- W2854323797 title "Particulate Progesterone-Binding Sites in the Equine Corpus Luteum1" @default.
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- W2854323797 doi "https://doi.org/10.1093/biolreprod/52.monograph_series1.419" @default.
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