Enhanced breakdown of arylsulfatase A in multiple sulfatase deficiency.
Enhanced breakdown of arylsulfatase A in multiple sulfatase deficiency.
复制标题
多种硫酸酯酶缺乏症中芳基硫酸酯酶 A 的分解增强。
DOI：
10.1111/j.1432-1033.1982.tb19770.x
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
K. von Figura
中科院分区：
文献类型：
--
作者：
A. Waheed;A. Hasilik;K. von Figura
多种硫酸酯酶缺乏症（粘液硫酸酯酶缺乏症）是一种溶酶体贮积症，其特征是所有已知硫酸酯酶的活性降低。为了测量多种硫酸酯酶缺乏症中芳基硫酸酯酶 A 的表观合成速率和半衰期，对来自疾病患者和对照的成纤维细胞进行了放射性氨基酸的脉冲追踪标记。通过免疫沉淀从细胞和培养基中分离芳基硫酸酯酶 A 和组织蛋白酶 D（一种不受多种硫酸酯酶缺乏症影响的溶酶体酶）。通过聚丙烯酰胺凝胶电泳分离标记的多肽，通过荧光照相术可视化并通过液体闪烁计数进行定量。使用单同位素和双同位素技术发现，与组织蛋白酶 D 相比，在多种硫酸酯酶缺乏症中，与对照成纤维细胞相比，芳基硫酸酯酶 A 的表观合成速率低 2--5 倍，半衰期短 4--9 倍。在多种硫酸酯酶缺乏的成纤维细胞中，从人尿和牛试验中分离出的内吞芳基硫酸酯酶A的内吞速率和稳定性与异染性脑白质营养不良成纤维细胞中的相同。我们假设在正常细胞中存在影响硫酸酯酶稳定性的基因产物，并且多种硫酸酯酶缺乏是由于该基因的突变造成的。
Multiple sulfatase deficiency (mucosulfatidosis) is a lysosomal storage disorder characterized by the decrease in activities of all known sulfatases. To measure the apparent rate of synthesis and the half-life of arylsulfatase A in multiple sulfatase deficiency, fibroblasts from patients with the disease and from controls were subjected to pulse-chase labelling with radioactive amino acids. Arylsulfatase A and cathepsin D, a lysosomal enzyme that is not affected in multiple sulfatase deficiency, were isolated from cells and media by immunoprecipitation. The labelled polypeptides were separated by polyacrylamide gel electrophoresis, visualized by fluorography and quantified by liquid scintillation counting. Using single and double isotope techniques it was found that, as compared to cathepsin D, the apparent rate of synthesis of arylsulfatase A was 2--5 times lower and the half-life 4--9-times shorter in multiple sulfatase deficiency than in control fibroblasts. In multiple sulfatase deficiency fibroblasts the rates of endocytosis and the stabilities of endocytosed arylsulfatases A isolated from human urine and bovine tests were equal to those in metachromatic leucodystrophy fibroblasts. We postulate that in normal cells a gene product exists that affects the stability of sulfatases and that multiple sulfatase deficiency is due to a mutation in this gene.