Reprogramming the enzymatic assembly line for site-specific fucosylation
Reprogramming the enzymatic assembly line for site-specific fucosylation
复制标题
重新编程酶装配线以进行位点特异性岩藻糖基化
DOI：
10.1038/s41929-019-0281-z
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发表时间：
2019
期刊：
影响因子：
37.8
通讯作者：
Cao Hongzhi
中科院分区：
文献类型：
--
作者：
Ye Jinfeng;Xia Hui;Sun Na;Liu Chang-Cheng;Sheng Anran;Chi Lianli;Liu Xian-Wei;Gu Guofeng;Wang Shu-Qi;Zhao Jie;Wang Ping;Xiao Min;Wang Fengshan;Cao Hongzhi
岩藻糖基化糖决定簇是细胞表面糖复合物和分泌性非结合聚糖的共同组成部分，在许多生理和病理过程中起着关键作用。刘易斯抗原的生物合成涉及多种岩藻糖基转移酶，其以非位点特异性方式催化聚-N-乙酰乳糖胺碳水化合物骨架的岩藻糖基化，从而产生异质和不完全岩藻糖基化的刘易斯抗原区域异构体。在本研究中，使用α 2，6-唾液酸化模块将α 2，6-连接的唾液酸引入特定位点作为岩藻糖基化的保护基团，从而以位点特异性方式精确控制聚-N-乙酰乳糖胺聚糖的酶促岩藻糖基化。岩藻糖基化后，唾液酸保护基可以很容易地被唾液酸酶去除，以提供多种具有确定岩藻糖基化模式的岩藻糖基化聚-N-乙酰基乳糖胺聚糖。该重编程酶组装线的普遍适用性和稳健性在22种复合刘易斯抗原和嵌合组织-血型抗原的合成中得到了例证，所述抗原和嵌合组织-血型抗原具有用于构建10种不同糖苷键的总共10种酶模块。
Fucosylated carbohydrate determinants are common components of cell surface glycoconjugates and secreted unconjugated glycans, which play pivotal roles in many physiological and pathological processes. The biosynthesis of Lewis antigens involves multiple fucosyltransferases that catalyse the fucosylation of the poly-N-acetyllactosamine carbohydrate backbone in a non-site-specific manner and thus generate heterogeneous and incompletely fucosylated Lewis antigen regioisomers. In this study, an α2,6-sialylation module was used to introduce α2,6-linked sialic acid to specific sites as the protecting group against fucosylation, thus precisely controlling enzymatic fucosylation of poly-N-acetyllactosamine glycans in a site-specific manner. The sialic acid protecting group can be easily removed by sialidase after fucosylation to provide a variety of fucosylated poly-N-acetyllactosamine glycans with defined fucosylation patterns. The general applicability and robustness of this reprogrammed enzymatic assembly line was exemplified in the synthesis of 22 complex Lewis antigens and chimeric histo-blood group antigens with a total of 10 enzyme modules for the construction of 10 different glycosidic linkages.