BIOLOGY OF THE HUMAN GLYCOPHORIN BLOOD GROUP ANTIGENS

人类血型血型抗原的生物学

• 批准号: 3365277
• 负责人: STEVEN L SPITALNIK
• 金额: $ 16.33万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3365277
• 关键词: Plasmodium falciparum; SDS polyacrylamide gel electrophoresis; animal tissue; antigen antibody reaction; antigen receptors; antiserum; blood group antigens; blood transfusion; carbohydrate structure; cell line; cellular immunity; complementary DNA; egg /ovum; enzyme linked immunosorbent assay; erythrocytes; glycophorin; glycoprotein biosynthesis; glycosylation; human tissue; immunofluorescence technique; intracellular transport; malaria; membrane proteins; molecular biology; monoclonal antibody; mutant; oligosaccharides; polymerase chain reaction; protein sequence; protein transport; reagent /indicator; site directed mutagenesis; surface antigens; transfection; western blottings

Glycophorin A is the major glycoprotein on the human red blood cell surface and contains both N- and O-linked oligosaccharides. Although structurally well characterized, relatively little is known concerning either the biosynthesis of glycophorin A or the role of glycosylation in translocation and surface expression of this molecule. Glycophorin A is important in the practice of transfusion medicine since it carries several different human blood group antigens, and antibodies to these antigens can cause hemolytic transfusion reactions, hemolytic disease of the newborn, and autoimmune hemolytic anemia. However, there have been few studies examining the fine specificity of binding of human polyclonal or mouse monoclonal antibodies to this molecule. Glycophorin A is also of medical importance because it can serve as the red blood cell surface receptor for the invasion of Plasmodium falciparum malaria merozoites. Due to the difficulty in obtaining mutant glycophorin A molecules with defined variations in amino acid sequence and oligosaccharide structure, a detailed understanding of this red blood cell-parasite interaction is not yet available. The goals of the current proposal are to study the biology of the human blood group glycophorin antigens by: 1) determining the importance of the N- and O-linked oligosaccharides in intracellular trafficking and surface expression of glycophorin A, 2) examining the fine specificity of the interactions between glycophorin A and human and mouse antibodies, and 3) determining the peptide and carbohydrate portions of glycophorin A which are recognized by human malaria parasites. These goals will be achieved by creating a series of cell lines stably transfected with either wild type or variant glycophorin A cDNA. Mutant glycophorin A cDNAs will be constructed by site-directed mutagenesis using the polymerase chain reaction. By expressing these cDNAs in both normal Chinese hamster ovary fibroblasts and those with defects in glycosylation, it will be possible to create variant glycophorin A molecules which differ in both amino acid and carbohydrate sequence. Both the intact cell lines and glycophorin A proteins purified from the cell lines will then be used to address these three goals.

糖蛋白A是人红细胞表面上的主要糖蛋白 并包含N-和O连接的寡糖。 虽然在结构上 特征很好，关于 糖蛋白A的生物合成或糖基化在易位中的作用 和该分子的表面表达。 糖蛋白A在 输血医学实践，因为它带有几种不同的人 血型抗原和对这些抗原的抗体会引起溶血 输血反应，新生儿的溶血疾病和自身免疫性 溶血性贫血。 但是，很少有研究检查罚款 人多克隆或小鼠单克隆抗体结合的特异性 到这个分子。 糖蛋白A也具有医学上的重要性 可以用作红细胞表面受体的入侵 恶性疟原虫疟疾植物。 由于困难 获取具有氨基中定义变化的突变糖蛋白A分子 酸序列和寡糖结构，对 这种红细胞 - 寄生虫相互作用尚不可用。 当前建议的目标是研究人类的生物学 血型糖蛋白抗原作者： 1）确定在N中N-和O连接的寡糖的重要性 糖蛋白A的细胞内运输和表面表达 2）检查糖蛋白A之间相互作用的精细特异性 以及人类和小鼠抗体，以及 3）确定糖蛋白A的肽和碳水化合物部分 被人类疟疾寄生虫认可。 这些目标将通过创建一系列细胞系来实现 用野生型或变异糖蛋白A cDNA转染。 突变体 糖蛋白A将通过使用位置定向诱变构建cDNA 聚合酶链反应。 通过在两个正常的情况下表达这些cDNA 中国仓鼠卵巢成纤维细胞和糖基化缺陷的仓鼠， 可能会创建变异的糖蛋白A分子，而不同的分子 在氨基酸和碳水化合物序列中。 完整的细胞系 然后将使用从细胞系纯化的蛋白质的糖蛋白A 解决这三个目标。