Horie K, Yano T, Funakoshi I, Yamashina I
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Clin Chim Acta. 1988 Sep 30;177(1):41-8. doi: 10.1016/0009-8981(88)90305-1.
In previous papers we reported that nucleotide pyrophosphatase (NPPase) activities are markedly elevated in skin fibroblasts from patients with Lowe's syndrome. To reveal the molecular basis for this elevation, we investigated to see whether or not the activities are proportional to the amounts of enzyme protein. Determination of the amounts of NPPase proteins by a radioimmunoassay using monoclonal antibodies revealed that the amounts of NPPase proteins were proportional to the enzyme activities. The NPPase proteins from patients' and normal cells were indistinguishable, after SDS-polyacrylamide gel electrophoresis, on Western blot analysis with staining with polyclonal antibodies raised against denatured NPPase. These data suggested that the elevated NPPase activities in fibroblasts in this disorder resulted from increased amounts of the enzyme proteins.
在之前的论文中，我们报道了低磷性抗维生素D佝偻病患者皮肤成纤维细胞中的核苷酸焦磷酸酶（NPPase）活性显著升高。为了揭示这种升高的分子基础，我们研究了这些活性是否与酶蛋白的量成正比。使用单克隆抗体通过放射免疫测定法测定NPPase蛋白的量，结果显示NPPase蛋白的量与酶活性成正比。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，用针对变性NPPase产生的多克隆抗体进行蛋白质印迹分析，患者细胞和正常细胞中的NPPase蛋白没有区别。这些数据表明，这种疾病中，成纤维细胞中NPPase活性升高是由于酶蛋白量的增加。
