Site‐Specific Protein Modification with Reducing Carbohydrates
Site‐Specific Protein Modification with Reducing Carbohydrates
复制标题
减少碳水化合物的位点特异性蛋白质修饰
DOI：
10.1002/ange.202116545
复制
发表时间：
2022
影响因子：
--
通讯作者：
Weimin Xuan
中科院分区：
文献类型：
--
作者：
Qifan Wu;Weidong Dong;Hui Miao;Qian Wang;Suwei Dong;Weimin Xuan
蛋白质糖基化在许多生物过程中起着至关重要的作用。然而，天然糖蛋白中寡糖的异质性和构建人类设计的糖蛋白的困难阻碍了糖生物学的基础研究和应用。在这里，我们描述了一种半合成的方法来用还原碳水化合物来定点修饰蛋白质。该方法包括基因掺入侧链酯化的天冬氨酸，然后将其定量转化为丙氨酸-β-酰肼(Aβz)，以及Aβz与一系列容易获得的还原碳水化合物的化学选择性结合。由此得到的Aβz连接的GlcNAc是对天然N-GlcNAc的紧密模拟，并且可以安装在各种蛋白质上，包括IL-17A和RNaseA。值得注意的是，蛋白质上的Aβz连接的GlcNAc通过内切糖苷酶催化的转糖基化反应与二天线低聚糖恶唑啉衍生物反应，使蛋白质上能够组装均一的糖链。
Protein glycosylation plays critical roles in many biological processes. However, the fundamental study and application of glycobiology are hindered by the heterogeneousness of oligosaccharides in natural glycoproteins and the difficulty in constructing glycoproteins of human design. Herein, we describe a semisynthetic method to site‐specifically modify proteins with reducing carbohydrates. The method involves the genetic incorporation of a side‐chain‐esterified aspartate, which was subsequently quantitatively converted into alanine‐β‐hydrazide (Aβz), and chemoselective conjugation of Aβz with a range of readily available reducing carbohydrates. The resulting Aβz‐linked GlcNAc is a close mimic of native N‐GlcNAc and could be installed on various proteins, including IL‐17A and RNase A. Notably, Aβz‐linked GlcNAc on proteins reacted with biantennary oligosaccharide oxazoline derivatives through endoglycosidase‐catalyzed transglycosylation reactions to enable the assembly of homogeneous glycans on proteins.