Two-step derivatization and mass spectral distinction of α2,3 and α2,6 sialic acid linkages on N-glycans by MALDI-TOF

Two-step derivatization and mass spectral distinction of α2,3 and α2,6 sialic acid linkages on N-glycans by MALDI-TOF
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通过 MALDI-TOF 对 N-聚糖上的 α 2,3 和 α 2,6 唾液酸键进行两步衍生化和质谱区分

DOI:
10.1016/j.cclet.2018.12.016
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发表时间:
2019-03
影响因子:
9.1
通讯作者:
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中科院分区:
化学1区
文献类型:
--
作者:
周小满;杨上;杨刚龙;谭增琦;关锋

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唾液酸化是人类重要的糖基化反应之一,在肿瘤的发生发展中起着重要作用,α 2,3-连接和α 2,6-连接的唾液酸通常位于N-聚糖末端,具有不同的功能。采用二甲胺和氢氧化铵两步衍生化方法对唾液酸进行修饰,使其更容易在MALDI-TOF MS上检测到不同的唾液酸键。利用该衍生化方法检测了胎球蛋白和乳铁蛋白等蛋白质样品中N-聚糖上的特异性唾液酸键。对于复杂细胞样品,在缺氧环境诱导的A549细胞中观察到双触角和三触角N-聚糖上的α 2,3-连接和α 2,6-连接唾液酸增加。总之,我们的唾液酸的两步衍生化提供了一种简单而准确的方法来检测特定的连接N-聚糖与MALDI-TOF质谱仪。
Sialylation is one of important glycosylation in human beings and plays an important role in cancer development.α2,3-Linked andα2,6-linked sialic acids are normally observed on the end ofN-glycans and have different functions. Derivatization on sialic acid was designed to detect the different linkages by MALDI-TOF MS. In this study, a two-step derivatization by dimethylamine and ammonium hydroxide was improved to modify the sialic acid and made it easier to detect the different linkages of sialic acids on MALDI-TOF MS. Using this derivatization method, specific sialic acids linkages onN-glycans of protein samples such as fetuin and lactoferrin were detected. For complex cell samples, increasedα2,3-linked andα2,6-linked sialic acids on bi-antennary and tri-antennaryN-glycans were observed in A549 cells induced by hypoxia environment. Taken together, our two-step derivatization of sialic acids offers a simple and accurate way to detect specific linkages onN-glycans with MALDI-TOF mass spectrometer.
DOI: 10.1073/pnas.1608069113
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