Integrated analysis of natural glycans using a versatile pyrazolone-type heterobifunctional tag ANPMP
Integrated analysis of natural glycans using a versatile pyrazolone-type heterobifunctional tag ANPMP
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DOI：
10.1016/j.carbpol.2023.121617
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发表时间：
2023-12-06
影响因子：
11.2
通讯作者：
Huang，Linjuan
中科院分区：
文献类型：
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作者：
Liu，Yuxia;Yang，Tong;Huang，Linjuan
聚糖通过碳水化合物-蛋白质相互作用介导各种生物学过程，聚糖微阵列已成为理解这些机制不可或缺的工具。然而，由于缺乏从糖缀合物获得具有不同结构的天然聚糖文库的方便和通用的方法，功能性糖组学的进展受到阻碍。为了应对这一挑战，我们开发了一种综合方法，该方法能够一锅释放并同时捕获，分离，结构表征和N/O-聚糖的功能分析。使用这种方法，将糖缀合物与吡唑啉酮类异双功能标签-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、大豆蛋白、人乳寡糖等中制备了一个标记的聚糖库。在N-乙酰化和全甲基化衍生化后，通过ESI-MS分析对聚糖进行了详细的结构表征，显示了>83个含有各种天然聚糖表位的高纯度聚糖-AEPMP。鸟枪微阵列的构建，研究蛋白质和抗血清的聚糖结合的细节。
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 ofN/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 byN-acetylation and permethylation, glycans were subjected to detailed structural characterization by ESI-MSnanalysis, 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.