Department für Chemie, Universität für Bodenkultur, Wien, Austria.
Department für Chemie, Universität für Bodenkultur, Wien, Austria; Institut für Parasitologie, Veterinärmedizinische Universität Wien, Wien, Austria.
Mol Cell Proteomics. 2023 Mar;22(3):100505. doi: 10.1016/j.mcpro.2023.100505. Epub 2023 Jan 28.
Caenorhabditis elegans is a frequently employed genetic model organism and has been the object of a wide range of developmental, genetic, proteomic, and glycomic studies. Here, using an off-line MALDI-TOF-MS approach, we have analyzed the N-glycans of mixed embryos and liquid- or plate-grown L4 larvae. Of the over 200 different annotatable N-glycan structures, variations between the stages as well as the mode of cultivation were observed. While the embryonal N-glycome appears less complicated overall, the liquid- and plate-grown larvae differ especially in terms of methylation of bisecting fucose, α-galactosylation of mannose, and di-β-galactosylation of core α1,6-fucose. Furthermore, we analyzed the O-glycans by LC-electrospray ionization-MS following β-elimination; especially the embryonal O-glycomes included a set of phosphorylcholine-modified structures, previously not shown to exist in nematodes. However, the set of glycan structures cannot be clearly correlated with levels of glycosyltransferase transcripts in developmental RNA-Seq datasets, but there is an indication for coordinated expression of clusters of potential glycosylation-relevant genes. Thus, there are still questions to be answered in terms of how and why a simple nematode synthesizes such a diverse glycome.
秀丽隐杆线虫是一种常用的遗传模式生物，已经成为了广泛的发育、遗传、蛋白质组学和糖组学研究的对象。在这里，我们使用离线 MALDI-TOF-MS 方法分析了混合胚胎和液体或平板培养的 L4 幼虫的 N-聚糖。在超过 200 种不同可注释的 N-聚糖结构中，观察到了不同阶段之间以及培养方式的变化。虽然胚胎期的 N-聚糖组总体上看起来不那么复杂，但液体和平板培养的幼虫在双分支岩藻糖甲基化、甘露糖的 α-半乳糖基化以及核心α1,6-岩藻糖的二-β-半乳糖基化方面存在差异。此外，我们通过 LC-电喷雾电离-MS 进行 β-消除分析了 O-聚糖；特别是胚胎期的 O-聚糖组包括一组以前未在线虫中显示存在的磷酸胆碱修饰结构。然而，聚糖结构的集合不能与发育 RNA-Seq 数据集中糖基转移酶转录本的水平明确相关，但表明与潜在糖基化相关基因的簇的协调表达。因此，在简单的线虫如何以及为什么合成如此多样化的聚糖方面仍有一些问题需要回答。
