Signal Transduction of GPR56

GPR56的信号转导

• 批准号: 8291805
• 负责人: Lei Xu
• 金额: $ 29.36万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8291805
• 关键词: Adhesions; Arrestins; Biological; Biological Assay; Biological Process; Cell secretion; Cell surface; Cells; Characteristics; Cleaved cell; Co-Immunoprecipitations; Data; Disease; Enzyme-Linked Immunosorbent Assay; Event; Family; Future; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Guanosine Triphosphate Phosphohydrolases; Human Development; Investigation; Label; Length; Link; Mammals; Mediating; Melanoma Cell; Membrane; Mission; Modeling; Monomeric GTP-Binding Proteins; Mutagenesis; Mutation; National Institute of General Medical Sciences; Outcomes Research; Phosphorylation; Phosphorylation Site; Pilot Projects; Play; Proline; Protein Kinase; Protein Kinase C; Proteins; Public Health; Radiolabeled; Reading; Receptor Activation; Recombinants; Regulation; Reporter; Reporting; Research; Role; Series; Serine; Signal Transduction; System; Tail; Testing; Threonine; Transmembrane Domain; United States National Institutes of Health; Vascular Endothelial Growth Factors; Work; cdc42 GTP-Binding Protein; cell behavior; desensitization; extracellular; human disease; in vivo; member; mutant; radiotracer; receptor; receptor function

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to understand how a family of G protein-coupled receptors (GPCRs), called adhesion GPCRs, transduces signals. Adhesion GPCRs have been recognized in recent years as critical regulators of diverse biological processes, but the mechanisms of their regulation remain unclear. A unique feature of adhesion GPCRs is that they are typically cleaved into two fragments. How these fragments contribute to their functions is not known. Substantial preliminary data have showed that the two cleaved fragments (GPRN and GPRC) of GPR56, a member of adhesion GPCRs, interact to regulate downstream signaling events, including the secretion of the vascular endothelial growth factor (VEGF) and the activation of protein kinase C¿ (PKC¿) in melanoma cells. The goal of the proposed research is to use this system to further dissect the activation mechanisms of GPR56 and its signaling components. Three specific aims are proposed: 1) Investigate whether GPRN regulates activities of GPRC. ELISA, co-immunoprecipitation, cell surface labeling, and mutagenesis analyses will be performed. 2) Examine whether GPRC regulates PKC¿ activity through G proteins. Minigene construct expression, small GTPase activity assay, reporter assays, and PKC membrane translocation analyses will be performed. 3) Investigate the desensitization mechanisms of GPRC. Mutagenesis analyses, radiolabeling assay, and expression analyses of shRNAs and mutant arrestins will be performed. Taken together; the outcome of the research will help to establish a paradigm on the signal transduction of adhesion GPCRs, a group of receptors that play essential roles in human development and diseases. Thus the proposed research is highly relevant to the mission of the National Institute of General Medical Sciences of NIH. PUBLIC HEALTH RELEVANCE: The proposed research aims to build a frame-work for studying the functions of a group of poorly characterized but highly disease-related receptors. It is therefore highly relevant to public health.

描述（由适用提供）：拟议的研究的长期目标是了解G蛋白偶联受体（GPCR）的家族如何转换信号。近年来，粘附GPCR被认为是潜水生物学过程的关键调节剂，但其调节的机制尚不清楚。粘附GPCR的一个独特特征是它们通常被切成两个片段。这些片段如何促进其功能。大量初步数据表明，GPR56的两个分裂片段（GPRN和GPRC）是粘附GPCR的成员，以调节下游信号事件，包括血管内皮生长因子（VEGF）的分泌和蛋白质激酶C¿拟议的研究的目的是使用该系统进一步剖析GPR56及其信号成分的激活机制。提出了三个具体目标：1）研究GPRN是否调节GPRC的活动。将进行ELISA，共免疫沉淀，细胞表面标记和诱变分析。 2）检查GPRC是否通过G蛋白来调节PKC？的活性。将进行微型构建体表达，小的GTPase活性测定，报告基因测定和PKC膜易位分析。 3）研究GPRC的脱敏机制。将进行诱变分析，放射性标记测定法和shRNA和突变阻滞蛋白的表达分析。一起;该研究的结果将有助于建立关于粘合剂GPCR的信号转导的范式，粘合剂GPCR是一组在人类发育和疾病中起重要作用的受体。拟议的研究与美国国立卫生研究院一般医学科学研究所的使命高度相关。 公共卫生相关性：拟议的研究旨在建立一个框架工作，以研究一组特征性较差但与疾病相关的受体的功能。因此，它与公共卫生高度相关。