Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor
Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor
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DOI：
10.1038/s41467-020-15192-1
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发表时间：
2020-03-13
影响因子：
16.6
通讯作者：
Kato, Koichi
中科院分区：
文献类型：
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作者：
Yagi, Hirokazu;Yagi-Utsumi, Maho;Kato, Koichi
MCFD2和ERGIC-53是V因子和VIII因子联合缺乏的致病基因的产物，在早期分泌途径中形成货物受体复合物，负责这些凝血因子的细胞内运输。在这项研究中，我们利用核磁共振技术成功地从因子VIII中鉴定出一个mcfd2结合片段，该片段由10个氨基酸序列组成，可以增强其分泌。这促使我们研究将该序列连接到重组糖蛋白上对其分泌的可能影响。我们发现，仅用护照序列标记重组红细胞生成素就能显著提高其分泌水平。我们的发现不仅为凝血因子的细胞内转运及其遗传缺陷提供了分子基础，而且为提高生物制药兴趣的重组糖蛋白的产量提供了潜在的有用工具。
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.