Identification and characterization of an estrogen-inducible glycoprotein (uterine secretory protein-1) synthesized and secreted by rat uterine epithelial cells.
Identification and characterization of an estrogen-inducible glycoprotein (uterine secretory protein-1) synthesized and secreted by rat uterine epithelial cells.
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大鼠子宫上皮细胞合成和分泌的雌激素诱导糖蛋白（子宫分泌蛋白-1）的鉴定和表征。
DOI：
10.1210/endo-122-1-105
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发表时间：
1988
期刊：
影响因子：
4.8
通讯作者：
S. Shimizu
中科院分区：
文献类型：
--
作者：
A. Takeda;N. Takahashi;S. Shimizu
由大鼠子宫上皮细胞合成了一种雌激素诱导的97kDa分泌型糖蛋白，命名为子宫分泌蛋白(USP)-1。将[35S]蛋氨酸直接注入大鼠子宫腔内，标记子宫腔液(ULF)蛋白。在去卵巢的大鼠中，[35S]蛋氨酸掺入Ulf蛋白的作用可以忽略不计。而去卵巢大鼠皮下注射17β-雌二醇(E_2)后，2天后[~(35)S]蛋氨酸掺入显著增加，表明雌二醇可诱导Ulf蛋白的从头合成和分泌。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和荧光法对标记的URF蛋白进行分析，鉴定出6个雌激素诱导多肽(130、110、97、65、42和39 kDa)。其中4个多肽(110、65、42和39 kDa)被固定的大鼠血清蛋白抗体吸附，表明这些多肽在抗原性上与血清蛋白相似。未被抗体柱吸附的两个多肽(130和97 kDa)被认为是子宫特异性分泌蛋白。通过制备性十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和电洗脱法进一步纯化了97 kDa的USP-1蛋白。纯化的USP-1与多种凝集素的反应性和糖组分分析表明，USP-1具有与岩藻糖连接的二天线N-连接复合型碳水化合物链。制备了能特异性免疫沉淀[35S]蛋氨酸标记的USP-1的兔多克隆抗体。斑点印迹酶免疫分析显示，E2处理4d的UF含有78.3+/-24.1(+/-SE)微克USP-1/mg蛋白。免疫组织化学染色显示，该蛋白仅定位于雌激素(E_2和己烯雌酚)处理的子宫上皮细胞。通过斑点印迹酶免疫分析、[35S]蛋氨酸标记的URF蛋白的免疫沉淀和子宫组织的免疫组织化学染色，发现睾酮、孕酮和地塞米松不能诱导USP-1的合成和分泌。本研究首次揭示了雌激素可诱导大鼠子宫上皮细胞合成和分泌特异性分泌蛋白，可作为分析雌激素作用分子机制的有用标记物。
An estrogen-inducible 97-kilodalton (kDa) secretory glycoprotein, designated as uterine secretory protein (USP)-1, synthesized by rat uterine epithelial cells was identified and characterized. Uterine luminal fluid (ULF) proteins were labeled by direct administration of [35S]methionine into uterine lumen of rats. Incorporation of [35S]methionine into ULF proteins was negligible in ovariectomized rats. However, when 17 beta-estradiol (E2) was administered in the ovariectomized rat as sc paraffin pellets, a marked increase of [35S]methionine incorporation was noted after 2 days of treatment, showing that the de novo synthesis and secretion of ULF proteins were induced by E2. Six estrogen-inducible polypeptides (130, 110, 97, 65, 42, and 39 kDa) were identified in the analysis of the labeled ULF proteins by sodium dodecylsulfate-polyacrylamide gel electrophoresis and fluorography. Four of these polypeptides (110, 65, 42, and 39 kDa) were adsorbed by the immobilized antibody against rat serum proteins, indicating that these polypeptides are antigenically similar to serum proteins. Two polypeptides (130 and 97 kDa) not adsorbed by the antibody column were suggested to be uterine-specific secretory proteins. The 97-kDa protein (USP-1) was further purified by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis and electroelution. Reactivity of purified USP-1 to various lectins and carbohydrate composition analysis suggested that USP-1 possesses biantennary N-linked complex-type carbohydrate chain with fucose. Rabbit polyclonal antibody which can specifically immunoprecipitate [35S]methionine-labeled USP-1 was developed. Dot blot enzyme immunoassay showed that 4-day E2-treated ULF contains 78.3 +/- 24.1 (+/- SE) micrograms USP-1/mg protein. Immunohistochemical staining of rat uterine tissue showed that this protein localized only in the epithelial cells treated with estrogen (E2 and diethylstilbestrol). Testosterone, progesterone, and dexamethasone failed to induce synthesis and secretion of USP-1 as assessed by dot blot enzyme immunoassay, immunoprecipitation of [35S]methionine-labeled ULF proteins, and immunohistochemical staining of uterine tissue. The present result, for the first time, revealed that estrogen can induce synthesis and secretion of specific secretory protein which could be the useful marker to analyze the molecular mechanism of estrogen action in rat uterine epithelial cells.
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DOI：
10.1016/0167-4781(83)90024-6
发表时间：
1983
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
DonohueJr,TM;Barker,KL
通讯作者：
Barker,KL
大鼠子宫雌激素反应细胞的原代培养：孕激素受体和分泌蛋白的诱导。
影响因子：
4.8
作者：
Kassis,JA;Sakai,D;Walent,JH;Gorski,J
通讯作者：
Gorski,J
未成熟大鼠子宫中的差异抗雌激素作用：对雌激素受体具有高亲和力的羟基化抗雌激素的比较。
DOI：
10.1016/0022-4731(83)90147-4
发表时间：
1983
期刊：
Journal of steroid biochemistry
影响因子：
--
作者：
Jordan,VC;Gosden,B
通讯作者：
Gosden,B
兔子宫上皮增殖与子宫珠蛋白产生之间的时间关系。
影响因子：
3.6
作者：
Murai,JT;Conti,CJ;Gimenez-Conti,I;Orlicky,D;Gerschenson,LE
通讯作者：
Gerschenson,LE
未成熟大鼠子宫中蛋白质合成的雌激素调节：孕酮对体外孵化期间释放到培养基中的蛋白质的影响。
影响因子：
4.8
作者：
Wheeler,C;Komm,BS;Lyttle,CR
通讯作者：
Lyttle,CR