Abstract：
Glycans mediate various biological processes through carbohydrate-protein interactions, and glycan microarrays have become indispensable tools for understanding these mechanisms. However, advances in functional glycomics are hindered by the absence of convenient and universal methods for obtaining natural glycan libraries with diverse structures from glycoconjugates. To address this challenge, we have developed an integrative approach that enables one-pot release and simultaneously capture, separation, structural characterization, and functional analysis of N/O-glycans. Using this approach, glycoconjugates are incubated with a pyrazolone-type heterobifunctional tag-ANPMP to obtain glycan-2ANPMP conjugates, which are then converted to glycan-AEPMP conjugates. We prepared a tagged glycan library from porcine gastric mucin, soy protein, human milk oligosaccharides, etc. Following derivatization by N-acetylation and permethylation, glycans were subjected to detailed structural characterization by ESI-MSn analysis, which revealed >83 highly pure glycan-AEPMPs containing various natural glycan epitopes. A shotgun microarray is constructed to study the fine details of glycan-bindings by proteins and antisera.
摘要：
聚糖通过碳水化合物-蛋白质相互作用介导各种生物过程，和聚糖微阵列已经成为理解这些机制不可或缺的工具。然而,缺乏从糖缀合物中获得具有不同结构的天然聚糖文库的方便且通用的方法阻碍了功能性糖组学的进展。为了应对这一挑战，我们开发了一种综合方法，可以实现一罐释放和同时捕获，分离,结构表征，和N/O-聚糖的功能分析。使用这种方法，糖缀合物与吡唑啉酮型异双功能标签-ANPMP孵育以获得聚糖-2ANPMP缀合物，然后将其转化为聚糖-AEPMP缀合物。我们从猪胃粘蛋白中制备了一个标记的聚糖文库，大豆蛋白，人乳寡糖，等。在通过N-乙酰化和全甲基化进行衍生化之后，通过ESI-MSn分析对聚糖进行详细的结构表征，其揭示了>83个含有各种天然聚糖表位的高纯度聚糖-AEPMPs。构建了shot弹枪微阵列，以研究蛋白质和抗血清的聚糖结合的精细细节。
