Functional Characterization of FKBP52 Interactions with Steroid Hormone Receptors

FKBP52 与类固醇激素受体相互作用的功能表征

基本信息

批准号：
7895986
负责人：
Marc B Cox
金额：
$ 19.76万
依托单位：
UNIVERSITY OF TEXAS EL PASO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7895986
关键词：
Affect Affinity Affinity Labels Androgen Receptor Androgens Binding Binding Proteins Binding Sites Biological Assay Chemosensitization Client Complex Cysteine Data Defect Development Disease Glean Glucocorticoid Receptor Glucocorticoids Goals Hormones Immunophilins Knockout Mice Lead Libraries Ligand Binding Domain Link Malignant Neoplasms Malignant neoplasm of prostate Mammalian Cell Mammals Mass Spectrum Analysis Mediating Modeling Molecular Chaperones Molecular Conformation Mus Mutation Nature Pharmaceutical Preparations Phenotype Photoaffinity Labels Physiological Play Progesterone Progesterone Receptors Proline Proteins Receptor Mediated Signal Transduction Regulation Reporter Role Site Specificity Staging Steroid Receptors Structure System Tacrolimus Binding Proteins Testing Therapeutic Yeasts affinity labeling base crosslink design dimer inhibitor/antagonist insight knockout gene mutant novel receptor receptor function reproductive research study response small molecule steroid hormone receptor tacrolimus binding protein 4 therapeutic target

项目摘要

DESCRIPTION (provided by applicant): The Hsp90 binding protein FKBP52, through interactions with the steroid hormone receptors, plays important physiological and potentially pathological roles in mammals. FKBP52 specifically regulates the androgen, progesterone and glucocorticoid receptors. Thus, FKBP52 represents an attractive therapeutic target for the treatment of hormone-dependent cancers such as prostate cancer. We have made progress in understanding the regions of functional importance on FKBP52, but we still do not know where the FKBP52 interaction site is on the receptors. The main hypothesis to be tested in this proposal is that FKBP52 potentiation of receptor function occurs through direct contacts between the FKBP52 FK1 domain and the receptor ligand binding domain leading to an enhancement of hormone binding. The dynamic nature by which the receptors achieve their hormone bound conformations to which FKBP52 associates makes it impractical to use a simple purified protein system. Thus, we must use more sophisticated approaches to analyze direct interactions between the steroid receptors and associated cochaperones. Each of the specific aims detailed below are independent from each other, but are all aimed at proving the proposed interaction between the FKBP52 FK1 domain and the receptor LBD and targeting that interaction with small molecule inhibitors. The long term goal of this project is the development of novel drugs for the treatment of hormone-dependent cancers. Towards this goal our immediate specific aims are: Aim #1: Use a previously validated androgen receptor-mediated reporter assay in yeast to identify random mutations within the receptor that alter the receptor's response to the FKBP proteins. Aim #2: Use directed photo affinity labeling of a cysteine-lacking FKBP52 mutant in combination with cross linking/mass spectrometry to identify and characterize the FKBP52-receptor interaction site. Aim #3: Screen compound libraries for selective FKBP52 inhibitors and characterize the specificity of inhibition and FKBP52 binding sites. We have identified two selective FKBP52 inhibitors to date.

描述（由申请人提供）：Hsp90结合蛋白FKBP52通过与类固醇激素受体相互作用，在哺乳动物中发挥重要的生理和潜在病理作用。 FKBP52 特异性调节雄激素、孕激素和糖皮质激素受体。因此，FKBP52代表了治疗激素依赖性癌症（例如前列腺癌）的有吸引力的治疗靶点。我们在了解 FKBP52 上具有重要功能的区域方面取得了进展，但我们仍然不知道 FKBP52 相互作用位点在受体上的位置。该提案要测试的主要假设是，FKBP52 受体功能的增强是通过 FKBP52 FK1 结构域和受体配体结合结构域之间的直接接触而发生的，从而导致激素结合的增强。受体实现与 FKBP52 结合的激素结合构象的动态性质使得使用简单的纯化蛋白质系统变得不切实际。因此，我们必须使用更复杂的方法来分析类固醇受体和相关辅助伴侣之间的直接相互作用。下面详述的每个具体目标彼此独立，但均旨在证明 FKBP52 FK1 结构域和受体 LBD 之间的拟议相互作用，并针对与小分子抑制剂的相互作用。该项目的长期目标是开发治疗激素依赖性癌症的新药。为了实现这一目标，我们当前的具体目标是：目标#1：使用先前在酵母中验证过的雄激素受体介导的报告基因检测来识别受体内的随机突变，这些突变会改变受体对 FKBP 蛋白的反应。目标#2：使用缺乏半胱氨酸的 FKBP52 突变体的定向光亲和标记结合交联/质谱来识别和表征 FKBP52-受体相互作用位点。目标#3：筛选选择性 FKBP52 抑制剂的化合物库，并表征抑制和 FKBP52 结合位点的特异性。迄今为止，我们已经鉴定出两种选择性 FKBP52 抑制剂。