Chemical Synthesis of an Erythropoietin Glycoform Having a Triantennary N-Glycan: Significant Change of Biological Activity of Glycoprotein by Addition of a Small Molecular Weight Trisaccharide,Journal of the American Chemical Society

当前位置： X-MOL 学术 › J. Am. Chem. Soc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Chemical Synthesis of an Erythropoietin Glycoform Having a Triantennary N-Glycan: Significant Change of Biological Activity of Glycoprotein by Addition of a Small Molecular Weight Trisaccharide
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-11-24 , DOI: 10.1021/jacs.0c08719
Yuta Maki _{1,

2} , Ryo Okamoto _{1,

2} , Masayuki Izumi ₁ , Yasuhiro Kajihara _{1,

2}

Affiliation

The glycosylation of proteins contributes to the modulation of the structure and biological activity of glycoproteins. Asparagine-linked glycans (N-glycans) of glycoproteins naturally exhibit diverse antennary patterns, such as bi-, tri-, and tetra-antennary forms. However, there are no chemical or biological methods to obtain homogeneous glycoproteins via the intentional alteration of the antennary form of N-glycans. Thus, the functions of the individual antennary form of N-glycan at a molecular level remain unclear. Herein, we report the chemical synthesis of an erythropoietin (EPO) glycoform having a triantennary sialylglycan at position 83, as well as two biantennary sialylglycans at both positions 24 and 38. We demonstrated efficient liquid-phase condensation reactions to prepare a sialylglycopeptide having a triantennary N-glycan prepared by the addition of a Neu5Ac-α-2,6-Gal-β-1,4-GlcNAc element to the biantennary glycan under semisynthetic conditions. The molecular weight of the newly added antennary element was ∼3% of the EPO glycoform, and the introduced position was the most distant from the bioactive protein. However, in vivo assays using mice revealed that the additional antennary element at position 83 dramatically increased the hematopoietic activity compared to a commercially available native EPO. These unprecedented data clearly indicate that the antennary pattern of N-glycans inherently plays a critical role in the modulation of protein functions.

中文翻译：

具有三触角 N-聚糖的促红细胞生成素糖型的化学合成：通过添加小分子量三糖显着改变糖蛋白的生物活性

蛋白质的糖基化有助于调节糖蛋白的结构和生物活性。糖蛋白的天冬酰胺连接聚糖 (N-聚糖) 自然表现出不同的触角模式，例如双触角、三触角和四触角形式。然而，没有化学或生物方法通过有意改变 N-聚糖的触角形式来获得均质糖蛋白。因此，N-聚糖的单个触角形式在分子水平上的功能仍不清楚。在此，我们报告了一种促红细胞生成素 (EPO) 糖型的化学合成，该糖型在 83 位具有三触角唾液聚糖，以及在 24 和 38 位具有两个双触角唾液聚糖。我们展示了有效的液相缩合反应来制备具有三触角 N-聚糖的唾液酸糖肽，该三触角糖肽通过在半合成条件下向双触角聚糖中添加 Neu5Ac-α-2,6-Gal-β-1,4-GlcNAc 元素而制备。新加入的触角元件的分子量约为 EPO 糖型的 3%，且引入位置距生物活性蛋白最远。然而，使用小鼠的体内试验表明，与市售的天然 EPO 相比，83 位的额外天线元件显着增加了造血活性。这些前所未有的数据清楚地表明，N-聚糖的天线模式固有地在蛋白质功能的调节中起着关键作用。4-GlcNAc 元件在半合成条件下连接到双天线聚糖。新加入的触角元件的分子量约为 EPO 糖型的 3%，且引入位置距生物活性蛋白最远。然而，使用小鼠的体内试验表明，与市售的天然 EPO 相比，83 位的额外天线元件显着增加了造血活性。这些前所未有的数据清楚地表明，N-聚糖的天线模式固有地在蛋白质功能的调节中起着关键作用。4-GlcNAc 元件在半合成条件下连接到双天线聚糖。新加入的触角元件的分子量约为 EPO 糖型的 3%，且引入位置距生物活性蛋白最远。然而，使用小鼠的体内试验表明，与市售的天然 EPO 相比，83 位的额外天线元件显着增加了造血活性。这些前所未有的数据清楚地表明，N-聚糖的天线模式固有地在蛋白质功能的调节中起着关键作用。使用小鼠进行的体内试验表明，与市售的天然 EPO 相比，83 位的附加天线元件显着增加了造血活性。这些前所未有的数据清楚地表明，N-聚糖的天线模式固有地在蛋白质功能的调节中起着关键作用。使用小鼠进行的体内试验表明，与市售的天然 EPO 相比，83 位的附加天线元件显着增加了造血活性。这些前所未有的数据清楚地表明，N-聚糖的天线模式固有地在蛋白质功能的调节中起着关键作用。

更新日期：2020-11-24

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南