MECHANISM OF SEX STEROID BINDING TO MACROMOLECULES

性类固醇与大分子的结合机制

• 批准号: 3311916
• 负责人: FREDERICK L SWEET
• 金额: $ 13.01万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-01-01 至 1985-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3311916
• 关键词: affinity chromatography; chemical aggregate; chemical synthesis; corpus luteum; embryo /fetus; enzyme structure; erythrocytes; hormone analog; hormone regulation /control mechanism; progesterone; radiotracer; sex hormones; steroid hormone metabolism

This project will continue investigation of the molecular events that determine the mechanism of steroid hormone binding to specific macromolecular binding sites. New affinity laveling steroids will be synthesized for these studies. Their effect on pregnancy will be tested in rats and sheep toilluminate the connection between the mechanism of steroid binding to macromolecules and the mechanism of steroid hormone action. Ovine fetal erythrocy te 20 Alpha-hydroxysteroid oxidoreductase (20 Alpha-HSD) will be prepared in quantity according to our newly established method established method for isolating 20 Alpha-hydroxysteroid oxidoreductase (EC 1.1.1.149) from bovine fetal erythrocytes. Both ovine and bovine 20 Alpha-HSD will be reacted with new adenosine and progesterone derivatives containing 3H and 14C bromoacetate side chains to characterize the active site amino acid compositions. The nucleosides will be used to continue our labeling of the cofactor binding region, and the steroids will be used for the steroid binding region of the active sties. Antibodies against ovine 20 Alpha-HSD will be raised in rabbits and used to measure levels of ovine fetal 20 Alpha-HSD in erythrocytes throughout pregnancy in cannulated sheep. By obtaining a profile for fetal 20 Alpha-HSD levels during gestation and also inactivating the enzyme in vivo with appropriate affinity labeling steroids we hope to learn the role of 20 Alpha-HSD in pregnancy. Th same antibodies will be used to measure conformational changes in 20 Alpha-HSD accompanying binding or progesterone or progesterone-protein conjugates to the enzyme. We will continue experiments with 17Beta-estradiol-7Alpha-butanoic acid - (rat) albumin in vivo to learn the precise mechanism of its uterotropic activity. Particular attention will be focused on evaluating the ability of the estrogen-protein conjugate to cross the target cell membrane and enter the nucleus. Definition of the mechanisms of steroid hormone binding to macromolecular binding sites of enzymes and receptor protein systems is the central goal of the project. This will braoden and deepen our understanding of the relationship between steriod structure and hormone action, leading to the design of new steroids for control of the mammalian reproductive system.

该项目将继续调查分子事件， 确定类固醇激素结合特异性 大分子结合位点。 新的亲和力laveling类固醇将 为这些研究而合成。 它们对怀孕的影响将在 大鼠和绵羊阐明类固醇的机制之间的联系， 与大分子的结合以及类固醇激素的作用机制。 绵羊胎儿红细胞20 α-羟基类固醇氧化还原酶（20 α-HSD）将根据我们新建立的 方法建立了分离20 α-羟基类固醇的方法 氧化还原酶（EC 1.1.1.149）。 两种绵羊 牛20 α-HSD将与新的腺苷和孕酮反应 含有3 H和14 C溴乙酸酯侧链的衍生物， 活性位点氨基酸组成。 核苷将用于 继续我们对辅因子结合区的标记，类固醇将 用于活性STIES的类固醇结合区。 抗体 将在家兔中饲养20只α-HSD，并用于测量 在整个妊娠期间， 插管羊 通过获得胎儿20 α-HSD水平的曲线， 在妊娠期间，也用适当的 亲和标记类固醇，我们希望了解20 α-HSD在 怀孕 相同的抗体将用于测量构象 伴随结合或孕酮的20 α-HSD的变化， 与酶结合的胆甾酮-蛋白质。 我们将继续 17 β-雌二醇-7 α-丁酸-（大鼠）白蛋白实验 体内研究其促子宫活性的确切机制。 将特别注意评价秘书处的能力， 雌激素-蛋白质缀合物穿过靶细胞膜并进入靶细胞。 原子核 类固醇激素结合的机制的定义 酶和受体蛋白系统的大分子结合位点是 项目的核心目标。 这将加深我们的 了解甾体结构与激素的关系 作用，导致设计新的类固醇控制哺乳动物 生殖系统