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劳氏肉瘤病毒转化的幼仓鼠肾细胞比幼仓鼠肾细胞表达更高水平的含有[GlcNAc-β(1,6)Man-α(1,6)Man]和多聚-N-乙酰乳糖胺序列的天冬酰胺连接的三触角和四触角糖肽。

Rous sarcoma virus-transformed baby hamster kidney cells express higher levels of asparagine-linked tri- and tetraantennary glycopeptides containing [GlcNAc-beta (1,6)Man-alpha (1,6)Man] and poly-N-acetyllactosamine sequences than baby hamster kidney cells.

作者信息

Pierce M, Arango J

出版信息

J Biol Chem. 1986 Aug 15;261(23):10772-7.

PMID:3015940
Abstract

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.

摘要

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。

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