Biological strategy for high quality and productivity of recombinant proteins in animal cells by the use of metal transporters

哺乳动物细胞用于生产重组蛋白，其翻译后修饰对其治疗功效是重要的。重组蛋白的生产由于需要新生多肽的高水平表达和适当加工而变得复杂，特别是在糖蛋白中，因为分泌途径中糖基化的能力在细胞中是有限的。在这项研究中，我们试图通过鉴定金属转运蛋白来改进生产重组糖蛋白的策略，所述金属转运蛋白在分泌途径中向糖基转移酶提供金属，例如锌和锰，因为这些金属作为酶的辅因子起作用。我们发现了Pmr1的一个同源物，它在酵母中具有钙通道的功能，对锰的转运很重要，并且可能在分泌途径中的糖基化反应中起作用，这可能导致重组细胞分泌能力的提高。

Mammalian cells are used for producing recombinant proteins whose post-translational modifications are important for their therapeutic efficacy. The manufacture of recombinant proteins is complicated by the need for both high levels of expression and appropriate processing of the nascent polypeptide, especially in glycoproteins, because the capacity of glycosylation in the secretory pathway is limited in the cells. In this study, we tried to improve the strategy for producing recombinant glycoproteins by identify metal transporters that supply metal such as zinc and manganese to glycosyltransferases in the secretory pathway because these metals are functional as cofactors of the enzymes. We found a homologue of Pmr1 that is a functional as calcium cannel in yeast is important for manganese transport and likely functional in glycosylation reaction in the secretory pathway, which may lead to the improvement of the secretion capacities of recombinant cells.