CHARACTERIZATION OF P60C-SRC BINDING TO MEMBRANES

P60C-SRC 与膜结合的表征

• 批准号: 3492443
• 负责人: COLIN GODDARD
• 金额: $ 4.99万
• 依托单位: OSI PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-03-01 至 1990-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3492443
• 关键词: binding proteins; cell membrane; chemical binding; colon neoplasms; erythrocyte membrane; membrane activity; membrane proteins; oncogenes; protein tyrosine kinase; tissue /cell culture

Human colon carcinoma cell lines have been shown by our own studies (S.I. Rayter, unpublished results) and by the report of Bolen et al., 1987 (1) to possess elevated levels of specific p60c-src protein-tyrosine kinase activity when compared to control colon lines. Two parameters appear to be essential for transformation by p60src; 1) the presence of an active protein tyrosine kinase and 2) the correct localization of the p60src protein at the plasma membrane. Recent studies have suggested the presence of a specific, high affinity p60src binding protein at the internal face of red cell ghost membranes which recognizes peptide sequences located within the 15 N-terminal residues of p60src (Goddard, manuscript submitted to J. Biol. Chem). The aims of this Phase I proposal are 1) to demonstrate specific high affinity binding of an N-terminal 19-mer p60src peptide to red cell ghost membranes and to colon carcinoma and normal colon plasma membranes, (ii) to demonstrate high affinity binding of purified human p60c-src to these membranes, and (iii) to cross-link purified p60c-src to the red cell membrane acceptor protein. In Phase II we propose to purify the specific plasma membrane p60c-src binding protein(s) from colon carcinoma and normal cell lines and develop assays for detection of agents enabling the therapeutic manipulation of this essential process in p60c-src mediated transformation.

我们自己的研究已表明人类结肠癌细胞系（S.I. Rayter，未发表的结果）和 Bolen 等人的报告，1987 (1) 特定 p60c-src 蛋白酪氨酸激酶水平升高 与对照结肠系相比的活性。 两个参数似乎是 对于 p60src 的转化至关重要； 1) 活性物质的存在 蛋白酪氨酸激酶和 2) p60src 的正确定位 质膜上的蛋白质。 最近的研究表明存在 特定的、高亲和力的 p60src 结合蛋白位于 红细胞鬼膜可识别位于其中的肽序列 p60src 的 15 个 N 末端残基（Goddard，手稿提交给 J. 生物。化学）。 第一阶段提案的目的是 1) 证明 N 端 19 聚体 p60src 肽的特异性高亲和力结合 红细胞鬼膜、结肠癌和正常结肠血浆 膜，(ii) 证明纯化的人的高亲和力结合 p60c-src 到这些膜上，并且 (iii) 将纯化的 p60c-src 交联到 红细胞膜受体蛋白。 在第二阶段，我们建议纯化 来自结肠的特定质膜 p60c-src 结合蛋白 癌细胞和正常细胞系并开发试剂检测方法 能够对 p60c-src 中的这一重要过程进行治疗操作 介导的转化。