ISOLATION AND CHARACTERIZATION OF PROGESTERONE RECEPTOR

孕酮受体的分离和表征

• 批准号: 6324700
• 负责人: DAVID O TOFT
• 金额: $ 15.73万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6324700
• 关键词: Saccharomyces cerevisiae; adenosine triphosphate; cell free system; chickens; gene deletion mutation; hormone binding protein; laboratory mouse; peptidylprolyl isomerase; progesterone receptors; protein structure function; receptor binding; reporter genes; reproductive hormone; reticulocytes; stress proteins

When in its inactive form in cytosol extracts, the avian progesterone receptor, like most steroid receptors, is complexed with the heat shock protein hsp90. This complex does not form through a simple association, but through a process that requires ATP hydrolysis and several additional proteins. other proteins in the receptor complex are hsp70 and three peptidylprolyl isomerases or immunophilins. Two of these, FKBP52 and FKBP54, bind the immunosuppressant drug FK506 and the third, CyP-40, binds cyclosporin A. A final protein in the receptor complex is p23 which is a unique phosphoprotein. We have made much progress in the description of receptor complexes and in the development of cell-free systems for their study. The objective of the proposed studies is to further our understanding on the assembly of receptor complexes, their dissociation upon hormone binding, and the functional significance of the receptor- associated proteins. This will be accomplished through the development and manipulation of chemically defined systems for receptor complex formation using purified proteins and specific antibody probes. Sub-complexes will also be analyzed to describe intermediates in complex formation and the various protein:protein interactions that occur. Finally, a yeast system with progesterone receptor will be used to identify the homologs of receptor-associated proteins in yeast and to test the biological significance of specific proteins of the complex by mutagenesis. These studies should provide new and valuable information on the earliest events in progesterone action and the axillary proteins that are needed for these events.

当在细胞质提取物中处于非活性形式时，禽类黄体酮 受体，像大多数类固醇受体一样，与热休克复合 蛋白质 hsp90。 这个复合体并不是通过简单的关联形成的， 但通过一个需要 ATP 水解和一些额外的过程 蛋白质。 受体复合物中的其他蛋白质是 hsp70 和三种 肽基脯氨酰异构酶或亲免素。 其中两个，FKBP52 和 FKBP54，结合免疫抑制剂药物 FK506，第三种，CyP-40，结合 环孢菌素 A。受体复合物中的最终蛋白质是 p23，它是 独特的磷蛋白。 我们在描述方面取得了很大进展 受体复合物及其无细胞系统的开发 学习。 拟议研究的目的是进一步推进我们的研究 了解受体复合物的组装及其解离 激素结合以及受体的功能意义- 相关蛋白质。 这将通过开发和 操纵化学确定的系统来形成受体复合物 使用纯化的蛋白质和特异性抗体探针。 子综合体将 也可以分析以描述复杂形成中的中间体和 发生的各种蛋白质：蛋白质相互作用。 最后，酵母系统 与孕酮受体的结合将用于鉴定 酵母中的受体相关蛋白并测试其生物学特性 通过诱变研究复合物特定蛋白质的重要​​性。 这些 研究应该提供有关最早事件的新的、有价值的信息 黄体酮作用以及这些作用所需的腋窝蛋白 事件。