MAMMALIAN CELL SURFACE GENETICS AND BIOCHEMISTRY

哺乳动物细胞表面遗传学和生物化学

• 批准号: 3276114
• 负责人: HENRY C WU
• 金额: $ 15.86万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-12-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3276114
• 关键词: acylation; affinity chromatography; autoradiography; carbohydrate structure; cell cell interaction; cell cycle; cell growth regulation; cell membrane; cell transformation; cytogenetics; electron microscopy; gel electrophoresis; gene expression; glycoproteins; hamsters; hybrid cells; immunodiffusion; lectin; membrane activity; membrane proteins; microfilaments; molecular genetics; protein biosynthesis; protein transport; radionuclides; temperature sensitive mutant; tissue /cell culture

We will continue to study the structure, functions, and dynamics of the surface membrane components of eukaryotic cells. We have used bacterial and plant toxins as probes to investigate the pathways for the entry and delivery of these toxins into the cytosol of Chinese hamster ovary (CHO) cells. We have employed these toxins to select somatic cell mutants defective in the intoxication process. Studies in the next five years will be directed towards (1) studies on the fatty acylation of membrane glycoproteins in CHO and yeast mutants altered in N- glycosylation; (2) isolation and characterization of CHO mutants resistant to ionophores monensin and nigericin; (3) isolation and characterization of Vero cell mutants defective in diphtheria toxin receptor; and (4) isolation and characterization of CHO mutants defective in the uptake, transport and release of toxins into the cytosol, including the conditionally lethal mutants. Techniques to be employed include molecular and somatic cell genetics and biochemical analysis of the structures, functions and dynamics of membrane components involved in the receptor- mediated endocytosis of toxins. Results from these studies should provide a better understanding of the intracellular protein transport process and the role of post-translational modification of membrane glycoproteins in cellular physiology.

我们将继续研究其结构、功能和动力学 真核细胞表面膜成分。 我们 使用细菌和植物毒素作为探针来研究 这些毒素进入和输送到体内的途径 中国仓鼠卵巢（CHO）细胞的细胞质。 我们已聘请 这些毒素可以选择有缺陷的体细胞突变体 中毒过程。 未来五年的研究工作将是 针对（1）膜脂肪酰化的研究 CHO和酵母突变体中的糖蛋白在N-中发生改变 糖基化； (2) CHO突变体的分离和表征 对离子载体莫能菌素和尼日利亚菌素具有抗性； (3)隔离和 白喉缺陷型 Vero 细胞突变体的表征 毒素受体； (4) CHO 的分离和表征 突变体在毒素的摄取、运输和释放方面存在缺陷 进入细胞质，包括条件致死突变体。 所采用的技术包括分子和体细胞 结构、功能和功能的遗传学和生化分析 参与受体的膜成分的动力学 介导的毒素内吞作用。 这些研究的结果应该 提供对细胞内蛋白质的更好理解 运输过程和翻译后修饰的作用 细胞生理学中膜糖蛋白的研究。