MURINE ASCITES TUMOR CELL GLYCOPROTEINS

鼠腹水肿瘤细胞糖蛋白

• 批准号: 3165294
• 负责人: IRWIN J GOLDSTEIN
• 金额: $ 19.57万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3165294
• 关键词: Ehrlich's tumor; alpha galactosidase; aminoacid analyzer; analytical ultracentrifugation; antibody; basement membrane; carbohydrate structure; cell membrane; disaccharides; electron microscopy; entactin; galactose; gel electrophoresis; immunofluorescence technique; laboratory mouse; laboratory rabbit; laminin; lectin; macrophage; malignant ascites; membrane proteins; protein biosynthesis; protein metabolism; protein sequence; protein structure; radiotracer; surface antigens; tissue /cell culture; tumor antigens

This research represents a continuation of our studies on alpha-D-galactosyl-containing glycoproteins present on certain animal cells and tissues. We will focus attention on the carbohydrate moieties of two basement membrane and cell surface glycoproteins: laminin and entactin, both of which contain alpha-D-galactosyl end groups. Laminin is believed to be present on the surface of highly metastatic cells. Structural and biosynthetic studies will be conducted on the carbohydrate residues of these glycoproteins as well as on the alpha-D-galactosyl-terminated glycoproteins present on the surface of Ehrlich tumor cells. Purified lectins of known carbohydrate binding specificity, antibodies raised against murine laminin and against a trisaccharide hapten present as part of the structure of these glycoproteins, will be employed in these studies. An attempt will be made to protect mice against a lethal dose of Ehrlich cells by immunizing the mice with the trisaccharide-protein conjugate. The appearance of cell surface laminin, which is expressed on stimulated but not resident murine macrophages, will be quantified under various conditions of stimulation using a lectin-enzyme conjugate. Biosynthetic studies will include an investigation of the incorporation of D-galactose and N-acetyl-D-glucosamine into the polylactosamineglycan that forms part of the structure of these alpha-D-galactosyl-terminated glycoproteins. We shall characterize an alpha-D-galactosidase present in Ehrlich ascites tumor cells and that probably is involved in the biodegradation of alpha-D-galactosyl-containing glycoproteins. The localization of alpha-D-galactosyl-terminated glycoconjugates in a variety of cells by electron microscopy will be conducted using a colloidal gold complex of Griffonia simplicifolia I-B4 isolectin, a probe specific for alpha-D-galactosyl end groups. We shall also attempt to select and characterize a variant of Ehrlich ascites tumor cells that lack alpha-D-galactosyl groups. (A)

这项研究代表了我们关于 某些动物细胞上存在的含α-D-D-半乳糖基糖蛋白 和组织。 我们将把注意力集中在两个的碳水化合物中 地下膜和细胞表面糖蛋白：层粘连蛋白和肠蛋白， 两者都包含α-D-半乳糖基端基。 相信层粘连蛋白 存在于高度转移细胞的表面上。 结构和 将对生物合成研究对碳水化合物残基进行 这些糖蛋白以及alpha-d-d-galactosyl终止 糖蛋白存在于Ehrlich肿瘤细胞的表面上。 纯化 已知碳水化合物结合特异性的凝集素，升高的抗体 反对鼠层粘连蛋白和针对三糖触觉的一部分 这些糖蛋白的结构将在这些结构中使用 研究。 将尝试保护小鼠免受致命剂量 ehrlich细胞通过用三糖蛋白免疫小鼠 共轭。 细胞表面层粘连蛋白的出现，该层蛋白在刺激上表达 但是，在各种 使用凝集素 - 酶结合物刺激条件。 生物合成研究将包括对掺入的研究 D-半乳糖和N-乙酰基-D-葡萄糖胺进入聚乳糖胺聚糖， 构成了这些α-D-半乳糖基终止的结构的一部分 糖蛋白。 我们将表征存在于 Ehrlich腹水肿瘤细胞，这可能与 含α-D-半乳糖基糖蛋白的生物降解。 Alpha-D-半乳糖基终止的糖缀合物的定位 电子显微镜将使用胶体进行各种细胞 格里富尼的黄金复合物I-B 4分离蛋白，特定于探针 用于α-D-半乳糖基端组。 我们还将尝试选择并表征Ehrlich的变体 缺乏α-D-半乳糖基群的腹水肿瘤细胞。 （一个）