Institute of Biochemistry, Department of Chemistry, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, Vienna, Austria.
Eric Kandel Institute, Department of Laboratory Medicine, Medical University of Vienna, Spitalgasse 23, Vienna, Austria.
Nat Commun. 2024 Nov 9;15(1):9725. doi: 10.1038/s41467-024-54134-z.
N-glycosylation is one of the most common protein modifications in eukaryotes, with immense importance at the molecular, cellular, and organismal level. Accurate and reliable N-glycan analysis is essential to obtain a systems-wide understanding of fundamental biological processes. Due to the structural complexity of glycans, their analysis is still highly challenging. Here we make publicly available a consistent N-glycome dataset of 20 different mouse tissues and demonstrate a multimodal data analysis workflow that allows for unprecedented depth and coverage of N-glycome features. This highly scalable, LC-MS/MS data-driven method integrates the automated identification of N-glycan spectra, the application of non-targeted N-glycome profiling strategies and the isomer-sensitive analysis of glycan structures. Our delineation of critical sub-structural determinants and glycan isomers across the mouse N-glycome uncovered tissue-specific glycosylation patterns, the expression of non-canonical N-glycan structures and highlights multiple layers of N-glycome complexity that derive from organ-specific regulations of glycobiological pathways.
N-糖基化是真核生物中最常见的蛋白质修饰之一，在分子、细胞和机体水平上都具有重要意义。准确可靠的 N-聚糖分析对于获得对基本生物学过程的系统理解至关重要。由于聚糖的结构复杂性，它们的分析仍然极具挑战性。在这里，我们公开了 20 种不同小鼠组织的一致 N-聚糖数据集，并展示了一种多模态数据分析工作流程，该工作流程能够以前所未有的深度和广度来研究 N-聚糖的特征。这种高度可扩展的、基于 LC-MS/MS 的、数据驱动的方法集成了 N-聚糖谱的自动识别、非靶向 N-聚糖分析策略的应用以及聚糖结构的异构体敏感分析。我们对小鼠 N-聚糖中关键亚结构决定因素和聚糖异构体的描绘揭示了组织特异性的糖基化模式、非典型 N-聚糖结构的表达，并强调了源自糖生物学途径的器官特异性调控的 N-聚糖复杂性的多个层次。
