SIGNAL TRANSDUCTION BY THE ECK RECEPTOR TYROSINE KINASE

ECK 受体酪氨酸激酶的信号转导

• 批准号: 2430280
• 负责人: VISHVA M DIXIT
• 金额: $ 21.43万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2430280
• 关键词: antibody; biological signal transduction; chemotaxis; enzyme activity; enzyme mechanism; gene expression; protein tyrosine kinase; receptor expression; site directed mutagenesis; yeasts

DESCRIPTION (Adapted from Investigator's Abstract): Receptor protein tyrosine kinases (RPTKs) are of fundamental importance in processes such as cell proliferation, survival, and differentiation. The Eph family is the largest subfamily of RPTKs with 13 members having been discovered to date. Eck was the first member of this subfamily for which a ligand was characterized. B61, a cytokine-inducible gene product was originally isolated from tumor necrosis factor-alpha stimulated endothelial cells. Endothelial cells were found to express the eck RPTK and it was further shown that B61 could act as an angiogenic factor in vivo and chemoattractant for endothelial cells in vitro. Using the yeast two-hybrid system, the applicant has identified two molecules that are involved in transducing signals from activated eck RPTK. The first was the p85 subunit of PI3-kinase. The second molecule identified was a novel gene product designated Src-like adapter protein (SLAP). The investigator has defined two Specific Aims. Specific Aim 1 is designed to determine the physiological consequence of activation of PI 3-kinase by the eck RPTK. This includes identifying the phosphotyrosine residue in the eck cytoplasmic domain that is engaged by the p85 subunit of PI-3 kinase. Additionally, by expressing dominant negative versions of the eck RPTK that are no longer able to activate PI-3 kinase, the investigator plans to determine the contribution of PI-3 kinase to known eck functions such as the induction of chemotaxis. Specific Aim 2 is designed to characterize SLAP, the first adapter molecule to be described in the src family. This will include defining the phosphortyrosine residue it binds to in the eck cytoplasmic domain and the downstream effector molecules it in turn engages using a combination of yeast 2 hybrid screening and traditional biochemical approaches. Further, a panel of monospecific antibodies will be raised against SLAP to determine its subcellular localization, phosphorylation status and whether it binds to the cytoplasmic domains of other cell surface receptors.

描述（改编自研究者摘要）：受体蛋白 酪氨酸激酶（RPTK）在诸如 如细胞增殖、存活和分化。以弗家族 是最大的 RPTK 亚家族，已发现 13 个成员 迄今为止。 Eck 是该亚家族的第一个配体成员 被表征。 B61，细胞因子诱导基因产物 最初是从肿瘤坏死因子-α刺激中分离出来的 内皮细胞。发现内皮细胞表达 eck RPTK 进一步表明B61可以作为血管生成因子 体内和体外内皮细胞的化学引诱剂。使用酵母 在双杂交系统中，申请人已经鉴定出两种分子 参与从激活的 eck RPTK 转导信号。第一个是 PI3 激酶的 p85 亚基。鉴定出的第二个分子是 新的基因产物被命名为 Src 样接头蛋白 (SLAP)。这 调查员定义了两个具体目标。设计具体目标1 确定 PI 3-激酶激活的生理后果 由 eck RPTK 提供。这包括鉴定磷酸酪氨酸残基 位于由 PI-3 的 p85 亚基参与的 eck 细胞质结构域中 激酶。此外，通过表达显性否定版本 eck RPTK 不再能够激活 PI-3 激酶， 研究人员计划确定 PI-3 激酶对已知的 eck 功能，例如诱导趋化性。具体目标 2 是 旨在表征 SLAP，第一个接头分子 在 src 系列中描述。这将包括定义 它与 eck 细胞质结构域中的磷酸酪氨酸残基结合，并且 它反过来使用组合来接合下游效应分子 酵母2杂交筛选和传统生化方法的研究。 此外，还将产生一组针对 SLAP 的单特异性抗体 确定其亚细胞定位、磷酸化状态和 是否与其他细胞表面的细胞质结构域结合 受体。