Synthesis and processing of arylsulfatase A in human skin fibroblasts.
Synthesis and processing of arylsulfatase A in human skin fibroblasts.
复制标题
人皮肤成纤维细胞中芳基硫酸酯酶 A 的合成和加工。
DOI：
10.1515/bchm2.1982.363.1.425
复制
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
通过在L-[4,5- 3h]亮氨酸或[2-3H]甘露糖中培养细胞，免疫沉淀法分离标记的芳基硫酸酯酶A，荧光图显示电泳分离的多肽，研究了正常和突变的人成纤维细胞中芳基硫酸酯酶A的生物合成。以芳基磺化酶A为前体合成，平均表观分子质量为62 kDa。在1 ~ 7天的追踪期内，细胞内前体转化为60.5 kDa的多肽。聚丙烯酰胺中60.5 kDa的产物与从人胎盘中分离的芳基磺化酶A中存在的两个多肽中的一个相对应。在异色性脑白质营养不良患者的成纤维细胞中未检测到可免疫沉淀的芳基磺化酶a多肽。在正常成纤维细胞中，不到10%的芳基硫酸酯酶A前体分泌到培养基中，而在粘脂病II型成纤维细胞和对照成纤维细胞中，高达90%的芳基硫酸酯酶A前体出现在培养基中，并在培养基中保持至少7天，而表观分子质量没有变化。芳基磺化酶A多肽似乎含有两个碳水化合物侧链。在大约90%的多肽中，两侧链都被内切- n -乙酰氨基葡萄糖酶H切割，而在其余的链中，两个低聚糖中的一个不被切割。
Biosynthesis of arylsulfatase A in normal and mutant human fibroblasts was studied by growing cells in the presence of L-[4,5-3H] leucine or [2-3H] mannose, isolation of labelled arylsulfatase A by immune precipitation and visualization of electrophoretically separated polypeptide by fluorography. Arylsulfatase A was synthesized as a precursor with a mean apparent molecular mass of 62 kDa. Intracellularly the precursor was converted into a 60.5 kDa polypeptide within a chase period of 1 to 7 days. The 60.5 kDa product in polyacrylamide corresponded to one of two polypeptides present in arylsulfatase A isolated from human placenta. In fibroblasts from a patient with metachromatic leukodystrophy no immune precipitable polypeptides of arylsulfatase A were detected. In normal fibroblasts less than 10% of the precursor of arylsulfatase A was secreted into the medium, whereas in mucolipidosis II fibroblasts and in control fibroblasts grown in the presence of NH4Cl up to 90% of the precursor of arylsulfatase A, appeared in the medium and remained there without change in the apparent molecular mass for at least 7 days. Arylsulfatase A polypeptides appear to contain two carbohydrate side chains. In about 90% of the polypeptides both side chains are cleaved by endo-beta-N-acetylglucosaminidase H, whereas in the remaining chains one of the two oligosaccharides is not cleaved.
亚基亲和层析及其在芳基硫酸酯酶 A 酶分离中的应用。
影响因子：
3.9
作者：
VanEtten,RL;Waheed,A
通讯作者：
Waheed,A