SURFACE OLIGOSACCHARIDES IN EMBRYONAL CARCINOMA CELLS

胚胎癌细胞中的表面寡糖

• 批准号: 3175420
• 负责人: RICHARD D CUMMINGS
• 金额: $ 11.22万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3175420
• 关键词: affinity chromatography; asparagine; biomedical equipment development; carbohydrate biosynthesis; carbohydrate sequence; carbohydrate structure; cell differentiation; cell membrane; chemical binding; embryo /fetus tissue /cell culture; endoderm; germ cell neoplasms; glycopeptides; glycosyltransferase; high performance liquid chromatography; immunochemistry; ion exchange chromatography; laboratory mouse; lectin; membrane proteins; neoplastic cell; neoplastic cell culture for noncancer research; oligosaccharides; oncoproteins; protein biosynthesis; protein structure; radionuclide double label; radiotracer; receptor; retinoate; tissue /cell culture; transferrin; tritium

Research demonstrated that differentiation of the mouse teratocarcinoma cell line F9 induced by retinoic acid is accompanied by major and specific alterations in the glycosylation of surface glycoproteins and suggest that the, changes may be due to modifications in activity of a few or perhaps a single glycosyltransferase. These studies do not, however, directly address many specific issues about the regulation of surface glycoprotein biosynthesis in animal cells. Little is actually known in detail about the structures of oligosaccharides at specific Asn-linked sugar sites of functional and normal cell surface glycoproteins (i.e. non- viral) and little is known about the factors that influence the glycosylation of surface proteins. Sugar addition is determined in part by the expression of appropriate glycosyltransferases an, glycosidases. However, several studies have shown that the primary sequences of protein can influence the glycosylation of the peptide at specific sites. To understand more about the factors regulating glycoprotein biosynthesis in F9 an, differentiated F9 cells, the glycosylation of a common surface glycoprotein, the transferrin receptor, will be investigated. The specific questions to be addressed in the proposed study are the following. (1) What are the structures of the chains at each Asn-linked glycosylation site on the receptor? (2) Does differentiation result in specific differences in the glycosylation of the receptor? (3) If changes are found to occur in the structures of the chain upon differentiation, how soon following the addition of retinoic acid are these change, observable? (4) Do the complex- type Asn-linked chains on the receptor have structures representative of the major types of chains derived from total glycoproteins? (5) If changes do occur in receptor glycosylation upon differentiation, do these changes affect the binding affinity of the receptor for transferrin or the turnover or recycling rate of the receptor? Answers to all these types of questions are not known for any surface glycoprotein in differentiating animal cells. Success in these studies will increase our knowledge of those factors influencing cell surface glycoprotein biosynthesis and should have broad implications for studies on other surface glycoproteins.

研究表明，小鼠的分化 视黄酸诱导的Teratocarcinoma细胞系F9 IS 伴随着糖基化的重大和特异性改变 表面糖蛋白的表面，并建议，可能是由于 修改几个或一个单一的活动 糖基转移酶。 但是，这些研究并不能直接解决许多具体的问题 有关表面糖蛋白生物合成的调节问题 在动物细胞中。 实际上，关于 特定ASN连接糖位点的寡糖结构 功能性和正常细胞表面糖蛋白（即非 - 病毒），对影响的因素知之甚少 表面蛋白的糖基化。 确定加糖 部分通过表达适当的糖基转移酶AN的表达 糖苷酶。 但是，一些研究表明主要 蛋白质序列可以影响 特定位置的肽。 更多地了解调节糖蛋白的因素 F9 AN分化F9细胞中的生物合成，糖基化 在常见的表面糖蛋白中，转铁蛋白受体将 被调查。 要解决的具体问题 拟议的研究是以下内容。 （1）每个ASN连接的链的结构是什么 受体上的糖基化位点？ （2）进行差异化 导致在糖基化的特定差异 受体？ （3）如果发现变化发生在结构中 差异后的链，加入后多久 视黄酸会发生变化吗？ （4） 受体上的类型ASN连接链具有结构 总体链的主要类型的代表 糖蛋白？ （5）如果受体糖基化确实发生了变化 分化后，这些变化会影响结合亲和力 转铁蛋白或周转或回收率的受体的 受体？ 所有这些类型的问题的答案不是 以分化动物细胞中任何表面糖蛋白而闻名。 这些研究的成功将增加我们对这些研究的了解 影响细胞表面糖蛋白生物合成和 应该对其他表面的研究具有广泛的影响 糖蛋白。