Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya, 467-8603, Japan.
Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, 444-8787, Japan.
Nat Commun. 2020 Mar 13;11(1):1368. doi: 10.1038/s41467-020-15192-1.
MCFD2 and ERGIC-53, which are the products of causative genes of combined factor V and factor VIII deficiency, form a cargo receptor complex responsible for intracellular transport of these coagulation factors in the early secretory pathway. In this study, using an NMR technique, we successfully identified an MCFD2-binding segment from factor VIII composed of a 10 amino acid sequence that enhances its secretion. This prompted us to examine possible effects of attaching this sequence to recombinant glycoproteins on their secretion. We found that the secretion level of recombinant erythropoietin was significantly increased simply by tagging it with the passport sequence. Our findings not only provide molecular basis for the intracellular trafficking of coagulation factors and their genetic deficiency but also offer a potentially useful tool for increasing the production yields of recombinant glycoproteins of biopharmaceutical interest.
MCFD2 和 ERGIC-53 是凝血因子 V 和 VIII 联合缺乏症的致病基因产物，它们形成一个货物受体复合物，负责这些凝血因子在早期分泌途径中的细胞内运输。在这项研究中，我们使用 NMR 技术成功地从因子 VIII 中鉴定出一个由 10 个氨基酸组成的 MCFD2 结合片段，该片段增强了因子 VIII 的分泌。这促使我们研究将这个序列连接到重组糖蛋白上是否会对它们的分泌产生可能的影响。我们发现，仅仅通过标记护照序列，重组促红细胞生成素的分泌水平就显著增加。我们的发现不仅为凝血因子的细胞内运输及其遗传缺陷提供了分子基础，而且为增加生物制药相关的重组糖蛋白的产量提供了一个潜在有用的工具。
