Construction of N-Glycan Microarrays by Using Modular Synthesis and On-Chip Nanoscale Enzymatic Glycosylation
Construction of N-Glycan Microarrays by Using Modular Synthesis and On-Chip Nanoscale Enzymatic Glycosylation
复制标题
DOI：
10.1002/chem.201001295
复制
发表时间：
2010-01-01
影响因子：
4.3
通讯作者：
Reichardt, Niels-Christian
中科院分区：
文献类型：
--
作者：
Serna, Sonia;Etxebarria, Juan;Reichardt, Niels-Christian
报道了一种有效的化学酶策略，该策略首次允许构建合成N聚糖的聚焦微阵列。各种N-聚糖核心结构已经被化学合成并共价固定在玻璃表面上。然后，通过放置在聚糖核心结构的各个点顶部的纳米液滴中的三种不同的糖基转移酶的作用，通过印刷机器人进行微阵列转化之后是对末端糖特异性的凝集素结合。这种纳米液滴中表面结合配体的酶促延伸相对于在溶液相中进行的反应将所需的宝贵糖基转移酶的量减少了7个数量级。此外，只有那些已经被证明是特异性糖基转移酶底物的配体才能被单独选择用于阵列上的延伸。结合聚焦模块化合成和纳米级芯片酶促延伸可以为急需的大合成glvcan阵列的快速构建开辟道路。
An effective chemoenzymatic strategy is reported that has allowed the construction, for the first time, of a focused microarray of synthetic N glycans Based on modular approaches, a variety of N-glycan core structures have been chemically synthesized and covalently immobilized on a glass sur face The printed structures were then enzymatically diversified by the action of three different glycosyltransferases in nanodroplets placed on top of individual spots of the microarray by a printing robot Conversion was followed by lectin binding specific for the terminal sugars This enzymatic extension of surface bound ligands in nano droplets reduces the amount of precious glycosyltransferases needed by seven orders of magnitude relative to reactions carried out in the solution phase Moreover, only those ligands that have been shown to be substrates to a specific glycosyltransferase can be individually chosen for elongation on the array The methodology described here combining focused modular synthesis and nanoscale on chip enzymatic elongation could open the way for the much needed rapid construction of large synthetic glvcan arrays