Hall C W, Robbins A R, Krag S S
Mol Cell Biochem. 1986 Nov-Dec;72(1-2):35-45. doi: 10.1007/BF00230634.
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[NAcGlcNH2]2-P-P-lipid, it fails to utilize these lipid intermediates to make normal amounts of [Glc]3[Man]9[NAcGlcNH2]2-P-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.
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早期步骤中发生改变的中国仓鼠卵巢细胞突变体。该程序可识别细胞内两种溶酶体水解酶（β-葡萄糖醛酸酶和α-艾杜糖醛酸酶）水平较低的细胞。通过这种方式分离出的一种突变细胞系CHB 11-1-3，与野生型细胞相比，其细胞内七种溶酶体酶的水平较低。尽管CHB 11-1-3能合成甘露糖基磷酸多萜醇和[Man]5[NAcGlcNH2]2-P-P-脂质，但它无法利用这些脂质中间体来合成正常量的[Glc]3[Man]9[NAcGlcNH2]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到溶酶体酶β-己糖胺酶上。此外，它的蛋白质糖基化不足。
