Evaluation of the context of downstream N- and free N-glycomic alterations induced by swainsonine in HepG2 cells
Evaluation of the context of downstream N- and free N-glycomic alterations induced by swainsonine in HepG2 cells
复制标题
评估 HepG2 细胞中苦马豆素诱导的下游 N- 和游离 N-糖组变化的背景
DOI：
10.1016/j.bbagen.2022.130168
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Shinohara Yasuro
中科院分区：
文献类型：
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作者：
Morikawa Chie;Sugiura Kanako;Kondo Keina;Yamamoto Yurie;Kojima Yuma;Ozawa Yurika;Yoshioka Hiroki;Miura Nobuaki;Piao Jinhua;Okada Kazue;Hanamatsu Hisatoshi;Tsuda Masumi;Tanaka Shinya;Furukawa Jun-ichi;Shinohara Yasuro
Swainsonine （SWA）是一种有效的II类α-甘露糖苷酶抑制剂，存在于世界各地的许多植物中，并在放牧这些植物的牲畜中引起严重的中毒。swa诱导动物中毒的机制尚不完全清楚。在本研究中，我们分析了向HepG2细胞中添加SWA后N-和游离N-糖偶联发生的改变，以更好地了解SWA诱导的糖偶联改变。在加入SWA后，我们观察到SWA特异性糖组化的出现，如独特的聚焦型杂交型和聚焦型M5 (M5F) n -聚糖，以及所有类别的Gn1 fng的显著增加。进一步分析这些糖化改变的背景表明，（聚焦的）杂化型n -聚糖不是这些Gn1 fng的前体，反之亦然。时间过程分析揭示了SWA暴露后糖偶联改变的动态性质，表明游离n -聚糖的积累早于杂交型n -聚糖的积累。杂交型n -聚糖，其中大多数是独特的核心聚焦，随着时间的推移，倾向于缓慢增加，正如M5F n -聚糖所观察到的那样。通过2-氟共加成抑制马豆素诱导的杂交n -聚糖和M5的独特聚焦，导致少西曼诺糖和聚焦的少西曼诺糖型n -聚糖以及少西曼诺糖型游离n -聚糖显著增加。结果不仅揭示了苦马豆素诱导HepG2细胞的总体糖糖化改变，而且为糖糖化改变之间的整体相互关系提供了信息。
Swainsonine (SWA), a potent inhibitor of class II α-mannosidases, is present in a number of plant species worldwide and causes severe toxicosis in livestock grazing these plants. The mechanisms underlying SWA-induced animal poisoning are not fully understood. In this study, we analyzed the alterations that occur in N- and free N-glycomic upon addition of SWA to HepG2 cells to understand better SWA-induced glycomic alterations. After SWA addition, we observed the appearance of SWA-specific glycomic alterations, such as unique fucosylated hybrid-type and fucosylated M5 (M5F) N-glycans, and a remarkable increase in all classes of Gn1 FNGs. Further analysis of the context of these glycomic alterations showed that (fucosylated) hybrid type N-glycans were not the precursors of these Gn1 FNGs and vice versa. Time course analysis revealed the dynamic nature of glycomic alterations upon exposure of SWA and suggested that accumulation of free N-glycans occurred earlier than that of hybrid-type N-glycans. Hybrid-type N-glycans, of which most were uniquely core fucosylated, tended to increase slowly over time, as was observed for M5F N-glycans. Inhibition of swainsonine-induced unique fucosylation of hybrid N-glycans and M5 by coaddition of 2-fluorofucose caused significant increases in paucimannose- and fucosylated paucimannose-type N-glycans, as well as paucimannose-type free N-glycans. The results not only revealed the gross glycomic alterations in HepG2 cells induced by swainsonine, but also provide information on the global interrelationships between glycomic alterations.