SURFACE MEMBRANES OF NORMAL AND CANCER CELLS

正常细胞和癌细胞的表面膜

• 批准号: 3165103
• 负责人: LEONARD WARREN
• 金额: $ 16.09万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-12-01 至 1991-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3165103
• 关键词: L cell; O glycosidase; affinity chromatography; carbohydrate structure; cell growth regulation; cell membrane; covalent bond; electrofocusing; enzyme mechanism; gene mutation; glycolipids; glycopeptides; glycoproteins; high performance liquid chromatography; human tissue; immunoglobulin G; laboratory mouse; laboratory rabbit; laboratory rat; mass spectrometry; membrane activity; membrane proteins; membrane structure; monoclonal antibody; neoplasm /cancer immunology; neoplastic cell; nuclear magnetic resonance spectroscopy; oncogenes; phagocytosis; phosphates; protein sequence; radionuclide double label; spectrometry; stoichiometry; temperature sensitive mutant; transferase; tritium

It has been found that incubation of UDP-galactose with GTP, Mn ions and extracts of transformed cells results in the formation of several rapidly migrating products. Two of these products are dominant, "Fast Peak 2" (FP2) and "Fast Peak 4" (FP4). The reaction is catalyzed by extracts of transformed cells with mutated or elevated levels of ras oncogene product (K-ras, H-ras and N-ras). Activity is also seen in human tumor cell lines (breast, lung, bladder), SV40-transformed cells and chemically transformed liver cells. The reaction is not detected in a variety of control cells, polyoma and Rous virus-transformed cells and some malignant cells. Preliminary experiments indicate that the reaction catalyzed by an extract of KMSV-transformed mouse fibroblasts can be inhibited by an anti-ras monoclonal antibody. Our aim is to: (a) obtain sufficient material to establish the structures of FP2 and FP4; (b) study their effects on cells and reactions related to growth control in order to establish their role in malignant (and normal) cells, i.e., whether the FP derivatives are involved in normal control of growth and differentiation; (c) fractionate the enzymes and factors involved in the synthesis of FP; (d) determine the nature of the reaction and the metabolic relation of FP2 and FP4; (e) establish that the ras gene product is a part of the mechanism and determine the action of the ras protein in normal and malignant cells; and (f) determine the distribution of the FP synthesizing enzymes in control and malignant tissues. We also plan to complete our studies in progress on: 1) the degradation of CMPNAN to form a putative CMPpyruvate. 2) comparison of glycopeptides of control and transformed cells. 3) the influence of environment and drugs on the carbohydrate component of IgG produced by hybridomas and the consequences of altered carbohydrate structure.

已发现UDP-半乳糖与GTP、Mn离子和 转化细胞的提取物导致几种快速的 迁移产品。 其中两款产品占据主导地位，“快峰2” (FP2)快速峰值4（Fast Peak 4） 该反应是催化的提取物， ras癌基因产物水平突变或升高的转化细胞 （K-ras、H-ras和N-ras）。 活性也见于人类肿瘤细胞系 （乳腺、肺、膀胱）、SV 40转化细胞和化学转化细胞 肝细胞 在多种对照细胞中未检测到该反应， 多瘤和Rous病毒转化的细胞和一些恶性细胞。 初步实验表明，由提取物催化的反应 的KMSV转化小鼠成纤维细胞可以被抗ras 单克隆抗体 我们的目标是：（a）获得足够的材料， （B）研究它们对细胞的作用和与 生长控制，以确定其在恶性（和正常） 细胞，即，FP衍生物是否参与正常控制 生长和分化;（c）分解酶和因子 参与FP的合成;（d）确定反应的性质 以及FP 2和FP 4的代谢关系;（e）确定ras基因 产品是机构的一部分，决定着RAS的作用 正常和恶性细胞中的蛋白质;和（f）确定分布 FP合成酶在正常组织和恶性组织中的表达。 我们也 计划完成我们正在进行的以下研究： 1)CMPNAN降解形成推定的CMP丙酮酸。 2)对照和转化细胞的糖肽的比较。 3)环境和药物对糖组成的影响 杂交瘤产生的IgG和碳水化合物改变的后果 结构