Ogilvie S, Buhi W C, Pawlina W, Larkin L H
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville 32610.
Biol Reprod. 1991 Dec;45(6):934-40. doi: 10.1095/biolreprod45.6.934.
This study was conducted to identify proteins synthesized and secreted de novo by the guinea pig uterus. Uterine samples were obtained from cycling, late-pregnant as well as ovariectomized and steroid-treated guinea pigs and cultured with either L-[3H]leucine or L-[35S]methionine. Two-dimensional SDS-PAGE of culture medium followed by fluorography was used to determine proteins synthesized and secreted de novo during a 24-h incubation period. Two complexes of estradiol-stimulated proteins (ESP) were detected. Each complex was composed of 5-7 unique proteins with slightly different isoelectric points. The higher molecular-weight complex had a molecular weight of 65,000-60,000 and an isoelectric point range of 5.2-6.1. The lower molecular-weight complex had a molecular weight of 60,000-55,000 and a similar range of isoelectric points. The two complexes of ESP were not observed in medium of explants from animals that received placebos, were late-pregnant, or were treated with progesterone only. Progesterone administered in combination with estradiol enhanced production of both complexes of ESP to similar degrees. Neither complex of ESP was secreted by the explant culture in the presence of tunicamycin, suggesting that the proteins are glycosylated. These findings demonstrate that the uterus of the guinea pig produces two unique complexes of proteins in response to estradiol stimulation, and all results are consistent with the hypothesis that ESP are contained in the carbohydrate-rich secretory granules of endometrial gland cells.
本研究旨在鉴定豚鼠子宫新合成并分泌的蛋白质。从处于发情周期、妊娠后期的豚鼠以及切除卵巢并用类固醇处理的豚鼠中获取子宫样本,并用L-[³H]亮氨酸或L-[³⁵S]甲硫氨酸进行培养。将培养基进行二维SDS-PAGE,然后进行放射自显影,以确定在24小时孵育期内新合成并分泌的蛋白质。检测到两种雌二醇刺激蛋白(ESP)复合物。每种复合物由5 - 7种等电点略有不同的独特蛋白质组成。分子量较高的复合物分子量为65,000 - 60,000,等电点范围为5.2 - 6.1。分子量较低的复合物分子量为60,000 - 55,000,等电点范围相似。在接受安慰剂、处于妊娠后期或仅用孕酮处理的动物的外植体培养基中未观察到这两种ESP复合物。与雌二醇联合使用的孕酮以相似程度增强了两种ESP复合物的产生。在衣霉素存在的情况下,外植体培养物未分泌任何一种ESP复合物,这表明这些蛋白质是糖基化的。这些发现表明,豚鼠子宫在雌二醇刺激下产生两种独特的蛋白质复合物,所有结果均与ESP包含在子宫内膜腺细胞富含碳水化合物的分泌颗粒中的假设一致。