BINDING AND FUNCTIONS OF RECEPTOR TYROSINE PHOSPHATASE B

受体酪氨酸磷酸酶 B 的结合和功能

• 批准号: 2272966
• 负责人: MARTIN H GRUMET
• 金额: $ 21.55万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2272966
• 关键词: antisense nucleic acid; blocking antibody; carbonate dehydratase; cerebellum; chemical binding; chick embryo; complementary DNA; developmental neurobiology; enzyme activity; fibronectins; gene mutation; glia; glioma; immunofluorescence technique; in situ hybridization; laboratory rat; neural cell adhesion molecules; oligonucleotides; protein sequence; protein tyrosine phosphatase; spinal cord; tenascin; tissue /cell culture; transfection

Cellular tyrosine phosphorylation plays a crucial role in the control of normal development and neoplasia. RPTP beta is a receptor protein tyrosine phosphatase that is expressed in glia in a pattern suggesting a role in morphogenesis and plasticity of the nervous system. Moreover, this protein binds to the extracellular matrix protein tenascin and to the neural cell adhesion molecules N-CAM and Ng. CAM/L1. The goal of this project is to analyze interactions of RPTP beta with various ligands and to study the consequences of these interactions on cell adhesion, and on transmembrane signalling that may modulate glial differentiation and interactions with neurons. The first specific aim is to characterize the expression patterns of RPTP beta in tissues and cells, and its interactions with different ligands during development. Then to analyze the binding properties of the different extracellular domains in RPTP beta cDNA constructs encoding different regions in RPTP beta will be used to express secreted and membrane-anchored forms of RPTP beta by transfection into mammalian cells. Molecular binding assays for secreted forms and cellular adhesion assays for forms expressed on the cell surface will be used to analyze which domains are important for binding of RPTP beta to different ligands such as tenascin and Ng-CAM/L1. To test potential functions of RPTP beta and its different domains in cells, molecular cloning techniques will be used to express or suppress expression of RFTP beta. In each case, effects of the treatment will be analyzed to detect changes in RPTP beta expression which will be correlated with alterations in cellular responses to ligands (i.e. tenascin and Ng-CAM) including cell adhesion, cell shape and cell division. RPTP beta is the first receptor/phosphatase with identified heterophilic ligands, and therefore it is important to investigate whether ligand binding to RPTP beta is involved in signal transduction by altering the specific activity of the phosphatase, by causing a redistribution of the phosphatase to alter its activity locally, or by changing the pattern of RPTP beta expression. This experimental approach will provide new information on the structure of binding regions of RPTP beta that may be involved in adhesion and growth regulation of normal and transformed astroglial cells. The results may also provide important clues for understanding development of radial glial cells and astrocytes, and their interactions with developing neurons, as well as potential roles of RPTP beta in growth of brain tumors.

细胞酪氨酸磷酸化在控制中起着至关重要的作用 正常发育和肿瘤。 RPTP Beta是一种受体蛋白 酪氨酸磷酸酶在胶质中以表明的模式表达 神经系统的形态发生和可塑性中的作用。而且， 该蛋白与细胞外基质蛋白替氏蛋白结合，并与 神经细胞粘附分子N-CAM和NG​​。 CAM/L1。目标的目标 项目是为了分析RPTP Beta与各种配体的相互作用， 研究这些相互作用对细胞粘附的后果以及对 可以调节神经胶质分化和的跨膜信号传导 与神经元的相互作用。第一个具体目的是表征 RPTPβ在组织和细胞中的表达模式，其表达模式 在开发过程中与不同配体的相互作用。然后分析 RPTP中不同细胞外域的结合特性 将使用编码RPTP Beta中不同区域的Beta cDNA构建体 表达分泌和膜的锚定形式的RPTP Beta。 转染到哺乳动物细胞。分泌的分子结合测定 在细胞上表达的形式的形式和细胞粘附测定 表面将用于分析哪些域对于结合很重要 RPTP Beta的不同配体，例如Tenascin和Ng-Cam/L1。测试 RPTPβ及其在细胞中不同域的潜在功能， 分子克隆技术将用于表达或抑制 RFTP Beta的表达。在每种情况下，治疗的影响都将是 分析以检测RPTPβ表达的变化，这将是 与细胞对配体的反应改变相关（即 Tenascin和ng-Cam）包括细胞粘附，细胞形状和细胞 分配。 RPTP Beta是第一个具有鉴定的受体/磷酸酶 异性配体，因此研究很重要 配体与RPTP Beta的结合是否参与信号转导 通过改变磷酸酶的特定活性，通过引起A 磷酸酶的重新分布以在局部改变其活性，或者通过 更改RPTPβ表达的模式。这种实验方法 将提供有关RPTP绑定区域结构的新信息 可能涉及正常和正常增长调节的β 转化的星形胶质细胞。结果也可能提供重要 了解径向神经胶质细胞和星形胶质细胞发展的线索， 他们与发展中神经元的互动以及潜力 RPTPβ在脑肿瘤生长中的作用。