GENETIC CONTROL OF PROTEOGYLCAN METABOLISM

蛋白聚糖代谢的遗传控制

• 批准号: 2176842
• 负责人: Jeffrey D Esko
• 金额: $ 21.67万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2176842
• 关键词: CHO cells; N acetylglucosamine; athymic mouse; carbohydrate biosynthesis; carbohydrate structure; cell adhesion; chondroitin sulfates; enzyme activity; enzyme deficiency; galactosyltransferases; glucuronosyltransferase; glycoprotein biosynthesis; glycosyltransferase; heparan sulfate; isomerase; molecular cloning; mutant; neoplastic transformation; phagocytosis; proteoglycan; sulfotransferase; transfection

The long range objective of this proposal is to gain insight into the mechanism of proteoglycan assembly. Altered proteoglycan synthesis correlates with the onset of several diseases, including neoplastic transformation, atherosclerosis, and various growth disorders associated with connective tissues. Thus, any insight into the mechanisms that cells use to synthesize and to regulate proteoglycan composition may provide new avenues for the development of therapeutic agents. To achieve these long-range goals, we propose a series of genetic, biochemical and molecular biologic experiments to probe the biosynthetic pathways of heparan sulfate and chondroitin sulfate synthesis in Chinese hamster ovary (CHO) cells. In particular we plan to execute the following studies: 1. Intermediates in Proteoglycan Biosynthesis. A comparison of proteo- glycans and biosynthetic intermediates in CHO mutants defective in chain initiation and polymerization may provide clues about the factors that control glycosaminoglycan composition. 2. Assembly of Heparan Sulfate. Studies of the reactions that lie at the branchpoint between the pathways of heparan sulfate and chondroitin sulfate synthesis may reveal how intermediates distribute into the individual pathways. Biochemical studies and genetic correction experiments of GlcNAc and GlcA transferase-deficient CHO cells should better define the relationship of these transferases. 3. Modification of Heparan Sulfate. Biochemical and genetic characterization of new mutants altered in bFGF binding should lead to a better understanding of polymer modification reactions involved in heparan sulfate synthesis. 4. Heparan Sulfate Structure and Biological Activity. Cell adhesion assays and binding experiments with heparin-binding proteins will help decide if unique saccharide sequences mediate the interaction of heparan sulfate with these ligands.

该提议的远距离目标是了解 蛋白聚糖组件的机理。 蛋白聚糖的合成改变 与几种疾病的发作相关，包括肿瘤 转化，动脉粥样硬化和相关的各种生长障碍 与结缔组织。 因此，任何对机制的见解 细胞用于合成和调节蛋白聚糖的组成可能 为开发治疗剂提供新的途径。 到 实现这些远程目标，我们提出了一系列遗传， 生化和分子生物学实验以探测生物合成 硫酸乙酰肝素和硫酸软骨素合成的途径 仓鼠卵巢（CHO）细胞。 特别是我们计划执行 以下研究： 1。蛋白聚糖生物合成中的中间体。 蛋白质的比较 在链中有缺陷的Cho突变体中的Glycans和生物合成中间体 启动和聚合可以提供有关因素的线索 控制糖胺聚糖组成。 2。硫酸乙酰肝素的组装。 对所在的反应的研究 硫酸乙酰肝素和软骨素的途径之间的分支 硫酸盐合成可能揭示了中间体如何分布到 单个途径。 生化研究和遗传矫正 GlcNAC和GlCA转移酶缺陷型CHO细胞的实验应 更好地定义这些转移酶的关系。 3。硫酸乙酰肝素的修饰。 生化和遗传 BFGF结合中改变的新突变体的表征应导致A 更好地了解涉及的聚合物修饰反应 硫酸乙酰肝素合成。 4。硫酸乙酰肝素结构和生物活性。 细胞粘附 用肝素结合蛋白进行测定和结合实验将有助于 确定独特的糖序序是否介导乙酰肝素的相互作用 用这些配体硫酸盐。