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锦龟输卵管中的雌激素结合蛋白:一种受体类型的证据。

Estrogen-binding proteins in the oviduct of the turtle, Chrysemys picta: evidence for a receptor species.

作者信息

Salhanick A R, Vito C C, Fox T O, Callard I P

出版信息

Endocrinology. 1979 Dec;105(6):1388-95. doi: 10.1210/endo-105-6-1388.

Abstract

Estradiol-binding proteins in the reproductive tract of the turtle, Chrysemys picta, were characterized. Cytosol was prepared from the oviducts of mature female turtles, and estradiol binding was measured using charcoal adsorption and glycerol density gradient centrifugation. A sex steroid-binding protein (SBP) similar to that found in turtle plasma was demonstrated in oviduct cytosol. The characteristics of this SBP-like binding were as follows: Ka = 10(8) M-1; capacity, 10(-12) mol/mg protein; and sedimentation coefficient, 6--7S in low salt gradients. The SBP-like protein binds testosterone and progesterone as well as 17 beta-estradiol but does not bind diethylstilbestrol. No receptor-like binding activity could be demonstrated using these techniques. Explant culture and DNA cellulose affinity chromatography were used to remove the SBP-like material before assay of [3H]estradiol binding. Using these techniques, a high affinity (Ka = 10(9) M-1), low capacity (n = 10(-14) mol/mg cytosol protein) estradiol receptor was demonstrated. The putative turtle receptor exhibits steroid specificity and sedimentation profiles (6S and 8S in low salt, 4S and 5S in high salt) comparable to those of estrogen receptors in mammalian species. These results suggest a certain degree of physiochemical similarity between putative estrogen receptors in mammalian and turtle reproductive tracts.

摘要

对彩龟(Chrysemys picta)生殖道中的雌二醇结合蛋白进行了表征。从成熟雌龟的输卵管制备胞质溶胶,并使用活性炭吸附和甘油密度梯度离心法测量雌二醇结合情况。在输卵管胞质溶胶中证实了一种与龟血浆中发现的类似的性类固醇结合蛋白(SBP)。这种类似SBP结合的特征如下:解离常数(Ka)= 10⁸ M⁻¹;结合容量,10⁻¹² mol/mg蛋白质;在低盐梯度中的沉降系数为6 - 7S。这种类似SBP的蛋白能结合睾酮、孕酮以及17β - 雌二醇,但不结合己烯雌酚。使用这些技术未证实有类似受体的结合活性。在测定[³H]雌二醇结合之前,采用外植体培养和DNA纤维素亲和色谱法去除类似SBP的物质。使用这些技术,证实了一种高亲和力(Ka = 10⁹ M⁻¹)、低容量(n = 10⁻¹⁴ mol/mg胞质溶胶蛋白)的雌二醇受体。推测的龟受体表现出类固醇特异性,其沉降图谱(低盐时为6S和8S,高盐时为4S和5S)与哺乳动物物种中的雌激素受体相当。这些结果表明哺乳动物和龟生殖道中推测的雌激素受体在一定程度上具有物理化学相似性。

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