OXIDASE-RELATED 48K PHOSPHOPROTEIN OF HUMAN NEUTROPHILS

人中性粒细胞的激活伴随着磷酸化 一种48K的磷蛋白。48K磷酸蛋白与蛋白质之间的联系 呼吸爆发氧化酶的激活过程是由 观察到这种蛋白质的磷酸化在两个方面是有缺陷的 各种形式的慢性肉芽肿疾病，一组遗传性疾病 在这种情况下呼吸爆发氧化酶无法发挥作用。我们建议 从分子水平和分子水平表征48K磷酸蛋白 它与其他氧化酶成分的相互作用。单克隆和多克隆 将获得针对48K磷酸蛋白的抗体并将其用于 用于分离和测序该蛋白质的cDNA克隆的探针。一次 这个克隆是可用的，它将被用作探测器来研究 该蛋白的染色体定位及其在细菌中的表达 吞噬细胞。将进行研究，以确定数量和地点 48K磷酸蛋白不同亚型上的磷酸基团。这个 磷蛋白将被提纯和结晶，以用于 确定了它的三维结构。最后，我们将检查 48K磷酸蛋白与其他氧化物组分的相互作用 用清洁剂激活无细胞系统中的氧化酶。

The activation of human neutrophils is accompanied by the phosphorylation of a 48K phosphoprotein. A connection between the 48K phosphoprotein and the process of respiratory burst oxidase activation is suggested by the observations that the phosphorylation of this protein is defective in two forms of chronic granulomatous disease, a group of inherited conditions in which the respiratory burst oxidase fails to function. We propose to characterize the 48K phosphoprotein in molecular terms and in terms of its interaction with other oxidase components. Monoclonal and polyclonal antibodies against the 48K phosphoprotein will be obtained and used as probes for isolating and sequencing a cDNA clone of the protein. Once this clone is available, it will be used as a probe to study the chromosomal location of the protein as well as its expression in phagocytes. Studies will be carried out to define number and location of phosphate groups on the various isoforms of the 48K phosphoprotein. The phosphoprotein will be purified and crystallized for purposes of determining its 3-dimensional structure. Finally, we will examine interactions of the 48K phosphoprotein with other oxidase components and with detergents that activate the oxidase in the cell-free system.