IMMUNOLOGICAL STUDIES OF THE DIGITALIS RECEPTOR

洋地黄受体的免疫学研究

• 批准号: 3343547
• 负责人: William James Ball
• 金额: $ 8.48万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-01 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3343547
• 关键词: adenosinetriphosphatase; antibody; antibody specificity; cell free system; chemical structure function; complementary DNA; digitalis; genetic translation; hybridomas; immunochemistry; immunopharmacology; kidney; messenger RNA; monoclonal antibody; ouabain

The membrane bound enzyme Na+,K+ ATPase is responsible for the active transport of Na+ and K+ across cell membranes and is, therefore, an essential component of all higher animal cells. It is also the putative receptor site for the cardiac glycosides which are specific inhibitors of its ATPase activity. This enzyme is of special interest because of its role in the maintenance of the electrophysiological properties of heart muscle cells. The goal of this project is to determine the membrane organization, structure, and function of the Na+, K+ ATPase and, thereby, aid in the elucidation of the mechanism of the inotropic effect of the cardiac glycosides. I plan to use antibodies directed against the "native" enzyme and its components in order to determine structure-function relationships. Studies using mouse monoclonal antibodies will investigate specific antibody-enzyme interactions in order to understand the mechanism(s) of antibody effects on enzyme function and the interactions between the different ligand binding sites of the enzyme. In addition, antibodies (polyclonal and monoclonal) with selective specificities towards the holoenzyme, the catalytic, glycoprotein subunit, and specific haptens will be used to determine the influence of the holoenzyme conformation on its antigenic properties and to identify, localize, and isolate functional regions of the enzyme containing the cardiac glycoside receptor, the phosphorylation site, and cation binding sites. This project will also provide the tools and develop the methodologies needed to study the biosynthesis and processing of the catalytic and glycoprotein subunits and to identify the cDNA or chromosomal clones of the catalytic subunit of the Na+, K+ ATPase.

膜结合酶Na+，K+ ATP酶负责活性 Na+和K+跨细胞膜的运输，因此， 所有高等动物细胞的基本组成部分。 这也是假定的 强心苷的受体位点，强心苷是 ATP酶活性。 这种酶特别令人感兴趣，因为它 维持心脏电生理特性的作用 肌肉细胞 本项目的目标是确定膜组织， 结构和功能的Na+，K+ ATP酶，从而帮助 心脏变力作用机制的阐明 糖苷 我计划使用针对“天然”酶的抗体 及其组分，以确定结构-功能关系。 使用小鼠单克隆抗体的研究将研究特异性 抗体-酶相互作用，以了解 抗体对酶功能的影响以及抗体与酶之间的相互作用 酶的不同配体结合位点。 此外，抗体 （多克隆和单克隆），具有针对 全酶、催化剂、糖蛋白亚基和特异性半抗原将 用于确定全酶构象对其构象的影响。 抗原特性，并鉴定、定位和分离功能性的， 含有强心苷受体的酶的区域， 磷酸化位点和阳离子结合位点。 该项目还将 提供必要的工具和方法， 催化和糖蛋白亚基的生物合成和加工， 为了鉴定该酶的催化亚基的cDNA或染色体克隆， Na+，K+ ATP酶。