Department of Chemistry, Yale University, New Haven, Connecticut, USA.
J Mass Spectrom. 2024 Jun;59(6):e5034. doi: 10.1002/jms.5034.
Glycosylation is an incredibly common and diverse post-translational modification that contributes widely to cellular health and disease. Mass spectrometry is the premier technique to study glycoproteins; however, glycoproteomics has lagged behind traditional proteomics due to the challenges associated with studying glycosylation. For instance, glycans dissociate by collision-based fragmentation, thus necessitating electron-based fragmentation for site-localization. The vast glycan heterogeneity leads to lower overall abundance of each glycopeptide, and often, ion suppression is observed. One of the biggest issues facing glycoproteomics is the lack of reliable software for analysis, which necessitates manual validation and serves as a massive bottleneck in data processing. Here, I will discuss each of these challenges and some ways in which the field is attempting to address them, along with perspectives on how I believe we should move forward.
糖基化是一种非常普遍和多样的翻译后修饰,广泛影响细胞的健康和疾病。质谱是研究糖蛋白的主要技术;然而,由于研究糖基化所带来的挑战,糖蛋白质组学落后于传统的蛋白质组学。例如,聚糖通过基于碰撞的碎片化而解离,因此需要基于电子的碎片化来进行位点定位。广泛的聚糖异质性导致每个糖肽的整体丰度降低,并且通常观察到离子抑制。糖蛋白质组学面临的最大问题之一是缺乏可靠的分析软件,这需要手动验证,并成为数据处理中的一个巨大瓶颈。在这里,我将讨论这些挑战中的每一个,以及该领域正在尝试解决这些挑战的一些方法,以及我认为我们应该如何向前发展的观点。