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Expression and functional characterization of a nucleotide sugar transporter from Drosophila melanogaster: relevance to protein glycosylation in insect cell expression systems

机译:果蝇的核苷酸糖转运蛋白的表达和功能表征:与昆虫细胞表达系统中的蛋白质糖基化有关。

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摘要

Insect cells are used routinely to express recombinant mammalian glycoproteins. However, insect protein glycosylation pathways are not well understood and appear to differ from those of mammalian cells. One way to more clearly evaluate the protein glycosylation potential of insect cells is to use the Drosophila melanogaster genome to identify genes that might encode relevant functions. These genes can then be expressed and the functions of the gene products directly evaluated by biochemical assays. In this study, we used this approach to determine the function of a putative Drosophila nucleotide sugar transporter gene. The results showed that this gene encodes a protein that can transport UDP-galactose, but not CMP-sialic acid. Thus, Drosophila encodes at least some of the infrastructure needed to produce glycoproteins with complex glycans, but this particular gene product does not directly support glycoprotein sialylation. These findings are relevant to insect cell biology and to an informed consideration of insect cell expression systems as tools for recombinant glycoprotein production. (C) 2002 Elsevier Science (USA). All rights reserved. [References: 60]
机译:昆虫细胞通常用于表达重组哺乳动物糖蛋白。但是,昆虫蛋白的糖基化途径尚未被很好地理解,并且似乎不同于哺乳动物细胞的途径。一种更清楚地评估昆虫细胞蛋白质糖基化潜力的方法是使用果蝇果蝇基因组来鉴定可能编码相关功能的基因。然后可以表达这些基因,并通过生化测定直接评估基因产物的功能。在这项研究中,我们使用这种方法来确定推定的果蝇核苷酸糖转运蛋白基因的功能。结果表明,该基因编码的蛋白质可以转运UDP-半乳糖,但不能转运CMP-唾液酸。因此,果蝇编码产生具有复杂聚糖的糖蛋白所需的至少一些基础设施,但是这种特定的基因产物不直接支持糖蛋白唾液酸化。这些发现与昆虫细胞生物学和昆虫细胞表达系统作为重组糖蛋白生产工具的知情考虑有关。 (C)2002 Elsevier Science(美国)。版权所有。 [参考:60]

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