Pierce M, Arango J
J Biol Chem. 1986 Aug 15;261(23):10772-7.
The alterations in complex-type N-linked oligosaccharides that can occur when an animal cell line is transformed by two dissimilar viruses were examined by comparing the N-linked oligosaccharides of baby hamster kidney (BHK) cells, metabolically radiolabeled with [2-3H]mannose, to the same class of oligosaccharides from BHK cells separately transformed by Rous sarcoma virus (RS-BHK), an RNA retrovirus, and polyoma virus (PY-BHK), a DNA papovavirus. Based on experiments that utilized serial lectin affinity chromatography, glycosidase digestions, and methylation analyses, both RS-BHK and PY-BHK cells demonstrated a significant increase in the relative amounts of tri- and tetraantennary complex-type N-linked oligosaccharides containing the branching sequence, [GlcNAc-beta(1,6)Man-alpha(1,6)Man], compared to the nontransformed BHK cells. In addition, almost all of the poly-N-acetyllactosamine sequence, [GlcNAc-beta(1,3)-Gal-beta(1,4)], was expressed on the tri- and tetraantennary N-linked oligosaccharides from BHK and RS-BHK cells that contain the sequence, [GlcNAc-beta(1,6)Man-beta(1,6)Man]. The increase in the relative amounts of this latter sequence in the transformed cells, therefore, most likely results in an increase in the amount of poly-N-acetyllactosamine sequence on the N-linked glycopeptides of these cells. The analysis of the degree of sialylation of the complex-type N-linked oligosaccharides from BHK and RS-BHK cells by ion exchange chromatography revealed no apparent differences, and in both of these cell types approximately 3% of the glycopeptide fraction radiolabeled with mannose was recovered in a highly negatively charged fraction that was identified by keratanase digestion to be keratan sulfate.
通过比较用[2-³H]甘露糖进行代谢性放射性标记的幼仓鼠肾(BHK)细胞的N-连接寡糖,与分别由RNA逆转录病毒劳氏肉瘤病毒(RS-BHK)和DNA乳头瘤病毒多瘤病毒(PY-BHK)转化的BHK细胞中同一类寡糖,研究了动物细胞系被两种不同病毒转化时复杂型N-连接寡糖可能发生的变化。基于利用系列凝集素亲和色谱、糖苷酶消化和甲基化分析的实验,与未转化的BHK细胞相比,RS-BHK和PY-BHK细胞中含有分支序列[GlcNAc-β(1,6)Man-α(1,6)Man]的三触角和四触角复杂型N-连接寡糖的相对量均显著增加。此外,几乎所有的多聚N-乙酰乳糖胺序列[GlcNAc-β(1,3)-Gal-β(1,4)]都表达在BHK和RS-BHK细胞中含有序列[GlcNAc-β(1,6)Man-β(1,6)Man]的三触角和四触角N-连接寡糖上。因此,转化细胞中后一种序列相对量的增加很可能导致这些细胞N-连接糖肽上多聚N-乙酰乳糖胺序列量的增加。通过离子交换色谱分析BHK和RS-BHK细胞中复杂型N-连接寡糖的唾液酸化程度,未发现明显差异,并且在这两种细胞类型中,用甘露糖放射性标记的糖肽部分中约3%在一个高度带负电荷的部分中回收,通过角蛋白酶消化鉴定该部分为硫酸角质素。