IMMUNOLOGICAL STUDIES OF THE DIGITALIS RECEPTOR

膜结合酶Na+，K+ATPase负责活性 Na+和K+跨细胞膜的运输，因此是一种 所有高等动物细胞的基本成分。它也是推定的 心脏糖苷的受体位置，这些糖苷是心脏的特异性抑制物 它的ATPase活性。这种酶特别令人感兴趣，因为它 在维持心脏电生理特性中的作用 肌肉细胞。 这个项目的目标是确定膜的组织结构， Na+，K+-ATPase的结构和功能，从而有助于 心脏变力作用机制的阐明 糖苷类药物。我计划使用针对“天然”酶的抗体。 及其组分，以便确定结构-功能关系。 使用小鼠单抗的研究将调查特定的 抗体-酶相互作用机制的研究(S) 抗体对酶功能的影响及其相互作用 酶的不同配体结合部位。此外，抗体 (多克隆和单克隆性)对 全酶、催化、糖蛋白亚基和特定的半抗原 用来测定全酶构象对其影响 抗原性及鉴定、定位和分离功能 含有心脏糖苷受体的酶的区域， 磷酸化部位和阳离子结合部位。该项目还将 提供研究所需的工具和方法 催化和糖蛋白亚基的生物合成和加工 目的：鉴定人乳头瘤病毒催化亚基的cdna或染色体克隆。 Na+，K+ATPase。

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.