LOCALIZATION OF LIGAND INTERACTIONS WITH THE SODIUM PUMP

描述：该项目的长期目标是了解 Na，K泵的结构-功能关系。调查员已经 蛋白质化学和亲和标记方面的拟议研究是 这是他以前工作的直接延伸。阳离子络合 泵的催化α亚基内的位置将由 带正电荷的羧基反应性化合物的亲和标记 (具体目标1)。主要成分中的异硫氰酸酯-活性赖氨酸 胞质结构域(Lys-501)将作为交联剂的锚 旨在识别与核苷酸有关的残基的实验 结合和能量转导(目标2)。额外的交联性和 光标记将用于直立的微粒子，以直接测试 根据水病分析提出的膜拓扑结构(目标3)。 最后，强心类固醇的光活性衍生物将用于 确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).