Aumiller Jared J, Jarvis Donald L
Department of Molecular Biology, University of Wyoming, P.O. Box 3944, Laramie, Wyoming 82071-3944, USA.
Protein Expr Purif. 2002 Dec;26(3):438-48. doi: 10.1016/s1046-5928(02)00550-8.
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.
昆虫细胞常被用于表达重组哺乳动物糖蛋白。然而,昆虫蛋白质糖基化途径尚未得到充分了解,且似乎与哺乳动物细胞的途径不同。更清楚地评估昆虫细胞蛋白质糖基化潜力的一种方法是利用黑腹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,并通过生化分析直接评估基因产物的功能。在本研究中,我们采用这种方法来确定一个推定的果蝇核苷酸糖转运蛋白基因的功能。结果表明,该基因编码一种能够转运UDP-半乳糖,但不能转运CMP-唾液酸的蛋白质。因此,果蝇编码了产生具有复杂聚糖的糖蛋白所需的至少一些基础结构,但这种特定的基因产物并不直接支持糖蛋白的唾液酸化。这些发现与昆虫细胞生物学以及将昆虫细胞表达系统作为重组糖蛋白生产工具的明智考量相关。