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通过外源性α2,8-Sialylation对原生细胞表面Sialoglycans的一步选择性标记

Jonathan L Babulic¹, Joshua M Kofsky², Marie E Boddington¹

¹Department of Biomedical and Molecular Sciences, Queen's University, Kingston K7L 3N6, Canada.

ACS chemical biology

|November 7, 2023

概括

这项研究引入了一种新型酶,Campylobacter jejuni sialyltransferase Cst-II I53S,用于修改具有特定糖结构的细胞表面. 这扩展了基于细胞的应用程序的糖基工程工具.

科学领域:

生物化学生化学
葡萄糖生物学葡萄糖生物学
细胞生物学细胞生物学

背景情况:

外酶性甘氨酸标签能够精确修改细胞表面的甘氨酸.
目前的方法为安装终端糖基因和糖基因表位提供了多功能工具.
这些修改对于调节细胞功能至关重要,并使得可选可视化或富含糖联物的可选可视化.

研究的目的:

确定和描述一种用于细胞表面甘氨酸修饰的新型酶.
扩大用于用α2,8-disialyl表位素进行外酶标记的工具包.
在本地细胞表面研究已识别的酶的特异性和有用性.

主要方法:

使用Campylobacter jejuni sialyltransferase Cst-II I53S进行糖修饰.
作为核酸糖探针,使用生物和亚酸标记的CMP-Neu5Ac衍生物.
将该方法应用于模型糖蛋白和各种细胞系的本地化细胞表面甘氨酸.

主要成果:

Cst-II I53S成功地将α2,8-disialyl表位物安装到细胞表面上.
修改后的甘氨酸在细胞表面停留至少6小时.
该酶主要标记为N-甘氨酸,有一些O-甘氨酸标记,并且显示出对α2,3-链接的酸的偏好.

结论:

Cst-II I53S是细胞表面甘氨酸工程酶的宝贵补充.
这种方法增强了对原生化甘氨酸的选择性标记.
这种方法适用于构建基于细胞的数组和推进糖生物学研究.

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