Preliminary characterization of a Chinese hamster ovary cell glycosylation mutant isolated by screening for low intracellular lysosomal enzyme activity.
Preliminary characterization of a Chinese hamster ovary cell glycosylation mutant isolated by screening for low intracellular lysosomal enzyme activity.
复制标题
通过筛选低细胞内溶酶体酶活性分离的中国仓鼠卵巢细胞糖基化突变体的初步表征。
DOI：
10.1007/bf00230634
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发表时间：
1986
影响因子：
4.3
通讯作者：
Krag,SS
中科院分区：
文献类型：
--
作者：
Hall,CW;Robbins,AR;Krag,SS
开发了一种新的筛选程序，用于分离在天冬酰胺连接糖蛋白生物合成的早期步骤中改变的中国仓鼠卵巢细胞突变体。该程序鉴定了两种溶酶体水解酶（β-葡萄糖醛酸酶和α-艾杜糖醛酸酶）细胞内水平较低的细胞。以这种方式分离的一种突变细胞系CHB 11-1-3与野生型细胞相比具有低的细胞内水平的七种溶酶体酶。虽然CHB 11-1-3合成甘露糖基磷酸多萜醇和[Man]5[NAcG 1cNH 2]2-P-P-脂质，但它不能利用这些脂质中间体来制备正常量的[Glc]3[Man]9[NAcG 1cNH 2] 2 P-P-脂质。由于这种糖基化缺陷，该突变体将与野生型不同结构的寡糖转移至溶酶体酶β-氨基己糖苷酶。此外，它使其蛋白质糖基化不足。
A novel screening procedure was developed for isolating Chinese hamster ovary cell mutants altered in the early steps of the biosynthesis of asparagine-linked glycoproteins. This procedure identifies cells with low intracellular levels of two lysosomal hydrolases, beta-glucuronidase and alpha-iduronidase. One mutant cell line isolated in this way, CHB 11-1-3, has low intracellular levels of seven lysosomal enzymes as compared to wild-type cells. Although CHB 11-1-3 synthesizes mannosylphosphoryldolichol and [Man]5[NAcG1cNH2]2-P-P-lipid, it fails to utilize these lipid intermediates to make normal amounts of [Glc]3[Man]9[NAcG1cNH2]2P-P-lipid. As a consequence of this glycosylation defect, this mutant transfers oligosaccharides of a different structure than wild type to the lysosomal enzyme beta-hexosaminidase. In addition, it underglycosylates its proteins.