Efficient Preparation and Bioactivity Evaluation of Glycan-Defined Glycoproteins,ACS Chemical Biology

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Efficient Preparation and Bioactivity Evaluation of Glycan-Defined Glycoproteins
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-11-24 , DOI: 10.1021/acschembio.0c00629
Ji Young Hyun _{1,

2} , Sanggil Kim ₃ , Chang-Hee Lee ₁ , Hyun Soo Lee ₃ , Injae Shin ₁

Affiliation

Owing to the generation of heterogeneous glycoproteins in cells, it is highly difficult to study glycoprotein-mediated biological events and to develop biomedical agents. Thus, general and efficient methods to prepare homogeneous glycoproteins are in high demand. Herein, we report a general method for the efficient preparation of homogeneous glycoproteins that utilizes a combination of genetic code expansion and chemoselective ligation techniques. In the protocol to produce glycan-defined glycoproteins, an alkyne tag-containing protein, generated by genetic encoding of an alkynylated unnatural amino acid, was quantitatively coupled via click chemistry to versatile azide-appended glycans. The glycoproteins produced by the present strategy were found to recognize mammalian cell-surface lectins and enter the cells through lectin-mediated internalization. Also, cell studies exhibited that the glycoprotein containing multiple mannose-6-phosphate residues enters diseased cells lacking specific lysosomal glycosidases by binding to the cell-surface M6P receptor, and subsequently migrates to lysosomes for efficient degradation of stored glycosphingolipids.

中文翻译：

聚糖定义的糖蛋白的有效制备和生物活性评估

由于细胞中异质糖蛋白的产生，研究糖蛋白介导的生物事件和开发生物医学制剂非常困难。因此，非常需要通用且有效的方法来制备均质糖蛋白。在此，我们报告了一种利用遗传密码扩展和化学选择性连接技术有效制备均质糖蛋白的通用方法。在生产聚糖定义的糖蛋白的协议中，通过对炔基化非天然氨基酸进行遗传编码生成的含炔标记的蛋白质通过点击化学定量耦合到多功能叠氮化物附加聚糖。发现由本策略产生的糖蛋白识别哺乳动物细胞表面凝集素并通过凝集素介导的内化进入细胞。此外，细胞研究表明，含有多个 6-磷酸甘露糖残基的糖蛋白通过与细胞表面 M6P 受体结合进入缺乏特定溶酶体糖苷酶的患病细胞，然后迁移到溶酶体以有效降解储存的鞘糖脂。

更新日期：2020-11-24

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