LOCALIZATION OF LIGAND INTERACTIONS WITH THE SODIUM PUMP

描述：本项目的长期目标是了解 Na，K泵的结构-功能关系。 研究者已 建议在蛋白质化学和亲和标记方面进行研究， 是他以前工作的直接延伸。 阳离子络合 泵的催化α亚基内的位点将通过以下方式来识别： 带正电荷的羧基反应性化合物的亲和标记 （具体目标1）。 异硫氰酸酯反应性赖氨酸主要 胞质结构域（Lys-501）将用作交联中的锚 旨在识别与核苷酸相关的残基的实验 结合和能量传递（Aim 2）。 额外的交联和 光标记将用于右侧外微粒体，以直接检测 提出的膜拓扑结构来自亲水性分析（目的3）。 最后，强心类固醇的光敏衍生物将用于 鉴定有助于糖苷结合的氨基酸（目标4）。

DESCRIPTION: The long-term goal of this project is an understanding of structure-function relationships in the Na,K-pump. The investigator has proposed studies in protein chemistry and affinity labeling that are a straightforward extension of his previous work. The cation complexation site within the catalytic alpha subunit of the pump will be identified by affinity labeling with positively charged carboxyl-reactive compounds (Specific Aim 1). The isothiocyanate-reactive lysine in the major cytoplasmic domain (Lys-501) will be used as an anchor in cross-linking experiments designed to identify those residues involved with nucleotide binding and energy transduction (Aim 2). Additional cross-linking and photolabeling will be used in right-side-out microsomes to test directly the proposed membrane topology derived from hydropathy analysis (Aim 3). Finally, photoactive derivatives of the cardiotonic steroids will be used to identify amino acids that contribute to glycoside binding (Aim 4).