Receptor-Signaling Protein Interactions in Real Time

受体信号蛋白实时相互作用

• 批准号: 7217980
• 负责人: SIDNEY PESTKA
• 金额: $ 33.97万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7217980
• 关键词: Address; Antibodies; Autoimmune Diseases; Biological Models; Cell physiology; Cell surface; Cells; Complex; Disease; Event; Fluorescence Resonance Energy Transfer; Goals; Human; Immune system; Immunoprecipitation; Interferon Gamma Receptor Complex; Interferon Type II; Interferon-alpha; Interferons; Kinetics; Lead; Life; Ligand Binding; Ligands; Malignant Neoplasms; Measurement; Measures; Mediating; Membrane; Methods; Mitosis; Movement; Phosphotransferases; Plasma; Play; Proteins; Receptor Signaling; Regulation; Research Personnel; Role; Screening procedure; Signal Transduction; Signaling Protein; Site; Structure; Technology; Time; Tissues; high throughput screening; immune function; programs; protein protein interaction; rapid technique; receptor

DESCRIPTION (provided by applicant): Interferon gamma (IFN-gamma) plays a crucial role in regulation of the immune system and is active on virtually all tissues. Its actions are mediated by its receptor that is the subject of this application. We previously identified two chains (IFN-gammaR1 and IFN-gammaR2) that are required for receptor activity. The current proposal focuses on the structure of the chains of the IFN-gamma receptor complex in live cells in the presence and absence of ligand. The specific aims directed to these goals are: to determine if all the IFN-gamma, receptor chains preassociated; the role of the Jak kinases in preassembly; determination of the sites on the receptor chains for preassembly; role of Stat1 in movement of the intracellular domains; determination of the kinetics of change in receptor structure after ligand binding; analysis of the kinetics and interactions of the downstream signaling components with the receptor complex; determine the sites of the receptor that serve as sites for cross-talk between the IFN-gamma and IFN-alpha receptor complexes. The configuration of the receptor chains will be studied by fluorescence resonance energy transfer in live cells. Because IFN-gamma plays a critical role in cellular and immune function, understanding all these aspects of its action will lead to better strategies to attack a variety of diseases such as cancers, and infectious and autoimmune diseases. Further application of this technology should lead to new rapid methods for high throughput screening and for identifying protein-protein interactions in live cells in real-time.

描述（由申请人提供）：干扰素γ (ifn - γ)在免疫系统的调节中起着至关重要的作用，几乎在所有组织中都有活性。它的作用是由受体介导的，受体是本应用的主题。我们之前确定了两条链（IFN-gammaR1和IFN-gammaR2）是受体活性所必需的。目前的建议集中在活细胞中存在和不存在配体的ifn - γ受体复合物链的结构。针对这些目标的具体目标是：确定所有ifn - γ受体链是否预先相关；Jak激酶在预组装中的作用；受体链上预组装位点的确定Stat1在胞内结构域运动中的作用；配体结合后受体结构变化动力学的测定下游信号组分与受体复合物的动力学和相互作用分析；确定受体的位置，作为ifn - γ和ifn - α受体复合物之间串扰的位置。受体链的结构将通过荧光共振能量转移在活细胞中进行研究。由于ifn - γ在细胞和免疫功能中起着至关重要的作用，了解其作用的所有这些方面将导致更好的策略来攻击各种疾病，如癌症，感染性和自身免疫性疾病。该技术的进一步应用将导致高通量筛选和实时鉴定活细胞中蛋白质-蛋白质相互作用的新的快速方法。