CYCLIC AMP BINDING PROTEINS AND RAS FUNCTION

环状 AMP 结合蛋白和 RAS 功能

• 批准号: 3503439
• 负责人: David Men-Hwei Tsai
• 金额: $ 5万
• 依托单位: SPHINX PHARMACEUTICALS CORPORATION
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3503439
• 关键词: adenosine diphosphate; adenosine monophosphate; adenosine triphosphate; animal tissue; binding proteins; bioassay; centrifugation; chemical binding; chromatography; cyclic AMP; cyclic GMP; drug screening /evaluation; immunoprecipitation; inhibitor /antagonist; microinjections; neutralizing antibody; protein purification; protooncogene; radiotracer; tissue /cell culture; transfection

Two CAMP binding proteins (50k and 10k daltons) that alter biochemical properties of protooncogene Ras in a CAMP dependent manner were isolated from bovine brain. In the presence of CAMP, the 50K dalton protein (p50) inhibited the nucleotide exchange reaction of Ras, whereas the 10K protein (p10) inhibited nucleotide binding. In the absence of cAMP, these two proteins are totally inactive. It is well established that protooncogene Ras plays an important role in controlling cell proliferation and differentiation. The interaction between these two proteins and Ras may represent a novel mechanism by which cell proliferation and differentiation are regulated. The objective of this phase I proposal is to partially purify these two proteins, established a standard assay and screen for the compounds or natural products that are able to alter the biochemical and biological activity of these two proteins. The compounds identified in this phase I study may develop into potential anti-proliferative drugs. In the phase II portion of this grant, these two proteins will be purified, sequenced and cloned. The biological significance of these two proteins will be elucidated by introducing these two proteins into cultured cells by protein microinjection and/or gene transfer techniques. Alternatively, antibodies that are able to neutralize these two proteins will be microinjected into cultured cells to determine the biological function of these two proteins. The cell model will be established to screen the compounds that are able to alter the biological activity of these two proteins.

两种改变生化的 CAMP 结合蛋白（50k 和 10k 道尔顿） 分离出 CAMP 依赖性方式的原癌基因 Ras 的特性 来自牛脑。 在 CAMP（50K 道尔顿蛋白 (p50)）存在的情况下 抑制Ras的核苷酸交换反应，而10K蛋白 (p10) 抑制核苷酸结合。 在没有 cAMP 的情况下，这两个 蛋白质完全不活跃。 众所周知，原癌基因 Ras 在控制细胞增殖和 差异化。 这两种蛋白和 Ras 之间的相互作用可能 代表了细胞增殖和分化的新机制 受到监管。 第一阶段提案的目标是部分 纯化这两种蛋白质，建立标准测定法并筛选 能够改变生物化学和化学性质的化合物或天然产物 这两种蛋白质的生物活性。 中鉴定的化合物 这项一期研究可能会开发成潜在的抗增殖药物。 在 在本次资助的第二阶段部分，这两种蛋白质将被纯化， 测序并克隆。 这两种蛋白质的生物学意义 将通过将这两种蛋白质引入培养细胞来阐明 蛋白质显微注射和/或基因转移技术。 或者， 能够中和这两种蛋白质的抗体将是 显微注射到培养细胞中以确定其生物学功能 这两种蛋白质。 建立细胞模型来筛选 能够改变这两种生物活性的化合物 蛋白质。