Glycosylation is one of the most complex post-translational protein modifications. Its importance has been established not only for eukaryotes but also for a variety of prokaryotic cellular processes, such as biofilm formation, motility and mating. However, comprehensive glycoproteomic analyses are largely missing in prokaryotes. Here we extend the phenotypic characterisation of N-glycosylation pathway mutants in Haloferax volcanii and provide a detailed glycoproteome for this model archaeon through the mass spectrometric analysis of intact glycopeptides. Using in-depth glycoproteomic datasets generated for the wild-type and mutant strains as well as a reanalysis of datasets within the Archaeal Proteome Project, we identify the largest archaeal glycoproteome described so far. We further show that different N-glycosylation pathways can modify the same glycosites under the same culture conditions. The extent and complexity of the Hfx. volcanii N-glycoproteome revealed here provides new insights into the roles of N-glycosylation in archaeal cell biology.

中文翻译：

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Haloferax volcanii的糖代谢组学揭示了广泛的糖蛋白组和不同N-糖基化途径的同时存在

糖基化是最复杂的翻译后蛋白质修饰之一。它的重要性不仅针对真核生物，还针对各种原核细胞过程，例如生物膜形成，运动性和交配，已经确立。但是，在原核生物中，缺少全面的糖蛋白组学分析。在这里，我们扩展了Haloferax volcanii中N-糖基化途径突变体的表型表征并通过完整糖肽的质谱分析为该模型古细菌提供详细的糖蛋白组学。使用为野生型和突变菌株生成的深入糖蛋白质组学数据集，以及对古细菌蛋白质组计划中数据集的重新分析，我们确定了迄今为止描述的最大的古细菌糖蛋白质组。我们进一步表明，不同的N-糖基化途径可以在相同的培养条件下修饰相同的糖位。Hfx的范围和复杂性。此处揭示的火山N-糖蛋白组学为N-糖基化在古细菌细胞生物学中的作用提供了新见解。