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一种雌激素依赖性猫子宫分泌蛋白(CUPED)的检测与纯化。

The detection and purification of a cat uterine secretory protein that is estrogen dependent (CUPED).

作者信息

Murray M K, Verhage H G, Buhi W C, Jaffe R C

出版信息

Biol Reprod. 1985 Jun;32(5):1219-27. doi: 10.1095/biolreprod32.5.1219.

Abstract

An estrogen-dependent secretory protein (CUPED) was detected and purified from uterine flushings of ovariectomized cats treated with 17 beta-estradiol. The protein was not detected in uterine flushings obtained from untreated ovariectomized animals or estrogen-primed animals treated with progesterone for 4 days. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of uterine flushings showed the presence of 1 or 2 protein bands with relative mobility values less than reduced and denatured thyroglobulin (Mr = 330,000). The protein was purified by differential centrifugation and gel filtration chromatography. Antiserum was raised against this purified protein in rabbits. The specificity of the antiserum to uterine fluid proteins was assessed by immunoblotting of electrophoretically transferred proteins. The antiserum cross-reacted with electrophoretically separated CUPED protein bands in uterine flushings. This protein may represent the content of the estradiol-induced secretory granules present in endometrial epithelial cells.

摘要

从用17β-雌二醇处理的去卵巢猫的子宫冲洗液中检测并纯化出一种雌激素依赖性分泌蛋白(CUPED)。在从未经处理的去卵巢动物或用孕酮处理4天的雌激素预处理动物获得的子宫冲洗液中未检测到该蛋白。子宫冲洗液的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示存在1条或2条蛋白带,其相对迁移率值小于还原和变性的甲状腺球蛋白(Mr = 330,000)。通过差速离心和凝胶过滤色谱法纯化该蛋白。用这种纯化蛋白在兔中制备抗血清。通过对电泳转移蛋白进行免疫印迹来评估抗血清对子宫液蛋白的特异性。该抗血清与子宫冲洗液中电泳分离的CUPED蛋白带发生交叉反应。这种蛋白可能代表子宫内膜上皮细胞中存在的雌二醇诱导的分泌颗粒的内容物。

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