Structural and Cellular Biology of GPH Receptors

GPH 受体的结构和细胞生物学

基本信息

批准号：
7541451
负责人：
JAMES Alan DIAS
金额：
$ 24.45万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-04-01 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7541451
关键词：
Affect Amino Acids Binding Cell Line Cell physiology Cell surface Cellular biology Complex Consensus Contraceptive methods Cytoplasmic Protein Defect Doctor of Philosophy Event Exhibits Female Fertility Follicle Stimulating Hormone Follicle Stimulating Hormone Receptor Funding G-Protein-Coupled Receptors Gametogenesis Glycoproteins Goals Gonadotropins Hormones Human Human Follicle Stimulating Hormone Link Literature Mediating Medical Ovary Pathway interactions Phosphorylation Play Preparation Principal Investigator Proteins Rattus Receptor Signaling Recycling Regulation Reproduction Role Signal Transduction Signal Transduction Pathway Site Small Interfering RNA Steroid biosynthesis Testis Thyroid Function Tests Ubiquitin Ubiquitination adapter protein casein kinase II desensitization difopein gonad function granulosa cell male mutant programs protein expression receptor receptor function sertoli cell tau Proteins trafficking

项目摘要

Receptors for the glycoprotein hormone follicle stimulating hormone (follitropin; FSH) are present in the Sertoli cells of testis and granulosa cells of ovaries. FSH is essential for normal high quality gametogenesis in males and females. Glycoprotein hormones which function in fertility and normal thyroid function are of general medical significance. Function of the FSH receptor (FSHR) beyond the initial binding event is regulated by cytoplasmic proteins which bind to receptor intracellular loops (il_) and regulate receptor function and act to organize receptors. Adapter proteins that have been identified which interact with human FSHR (hFSHR) are the focus of these studies. Aim one is to study the role of newly discovered adapter proteins APPL 1 and 2, that bind to FSHR-iLL Using siRNA to inhibit their synthesis in primary cultures of rat granulosa cells, the role of APPL1 and APPL2 regulation in FSH driven granulosa cell functions will be determined. In addition, amino acids in FSHR-IL1 which are essential for interaction with APPL proteins will be determined so that binding defective mutants can be studied for trafficking defects. Aim two will study the adapter protein 14-3- 3 tau which binds to FSHR-iL.2. It will be determined how inhibition of 14-3-3 tau affects FSHR signal transduction pathways in primary cultures of rat granulosa cells. Mutants ofhFSHR-il_2 defective in binding to 14-3-3 tau will be prepared and FSH activation of the Erk pathway will be studied in rat granulosa cell lines expressing these FSHR mutants. The third aim will determine the functional significance of the protein kinase CK2 (PKCK2) consensus site of thehFSHR-il_3. Consequences of inhibition of PKCK2 on steroidogenesis in primary cultures of human granulosa cells will be studied. It will be determined if PKCK2 interacts with hFSHR-il_3,and if consensus site mutants of the receptor are ubiquitination defective. Phosphorylation of hFSHR by PKCK2 will also be studied. The overall goal of the studies is to provide new information to better our understanding of regulation of gonadal function by gonadotropins and provide new avenues for fertility

存在糖蛋白激素卵泡刺激激素（follitropin; FSH）的受体在睾丸和卵巢颗粒细胞的塞托利细胞中。 FSH对于正常高质量至关重要男性和女性的配子发生。糖蛋白激素在生育和正常甲状腺功能具有一般的医学意义。 FSH受体（FSHR）以外的功能初始结合事件由细胞质蛋白调节，该蛋白与受体内环结合（IL_）并调节受体功能并起作用以组织受体。衔接蛋白已经确定哪些与人FSHR相互作用（HFSHR）是这些研究的重点。目标是研究新发现的新发现的衔接蛋白应用1和2的作用，该蛋白使用1和2与fshr-ill结合 siRNA抑制它们在大鼠颗粒细胞的原发性培养物中的合成，Appl1和将确定FSH驱动的颗粒细胞功能中的APPL2调节。另外，氨基酸在FSHR-IL1中，将确定与Appl蛋白相互作用至关重要的，以便结合可以研究有缺陷的突变体的贩运缺陷。目标两个将研究适配器蛋白14-3- 3 tau与fshr-il.2结合。将确定抑制14-3-3 TAU如何影响FSHR 大鼠颗粒细胞原发培养物中的信号转导途径。 HFSHR-IL_2的突变体将准备与14-3-3 Tau结合的缺陷，并且ERK途径的FSH激活将为在表达这些FSHR突变体的大鼠颗粒细胞系中进行了研究。第三个目标将决定蛋白激酶CK2（PKCK2）共有位点的功能意义。抑制PKCK2对人颗粒原代培养物中类固醇发生的后果细胞将被研究。如果PKCK2与HFSHR-IL_3相互作用，并且是否会确定共识网站受体的突变体泛素化缺陷。 PKCK2对HFSHR的磷酸化也将被研究。研究的总体目标是提供新信息，以更好地理解促性腺激素对性腺功能的调节，并提供新的生育途径