Department of Chemistry, Stanford University, Stanford, California, USA.
Department of Chemistry, Stanford University, Stanford, California, USA; Howard Hughes Medical Institute, Stanford, California, USA.
Mol Cell Proteomics. 2021;20:100029. doi: 10.1074/mcp.R120.002277. Epub 2020 Dec 20.
Glycosylation is a prevalent, yet heterogeneous modification with a broad range of implications in molecular biology. This heterogeneity precludes enrichment strategies that can be universally beneficial for all glycan classes. Thus, choice of enrichment strategy has profound implications on experimental outcomes. Here we review common enrichment strategies used in modern mass spectrometry-based glycoproteomic experiments, including lectins and other affinity chromatographies, hydrophilic interaction chromatography and its derivatives, porous graphitic carbon, reversible and irreversible chemical coupling strategies, and chemical biology tools that often leverage bioorthogonal handles. Interest in glycoproteomics continues to surge as mass spectrometry instrumentation and software improve, so this review aims to help equip researchers with the necessary information to choose appropriate enrichment strategies that best complement these efforts.
糖基化是一种普遍存在的、异质的修饰方式,在分子生物学中有广泛的影响。这种异质性排除了可以普遍有益于所有聚糖类的富集策略。因此,富集策略的选择对实验结果有深远的影响。在这里,我们综述了现代基于质谱的糖蛋白质组学实验中常用的富集策略,包括凝集素和其他亲和层析、亲水相互作用层析及其衍生物、多孔石墨碳、可逆和不可逆化学偶联策略,以及经常利用生物正交处理的化学生物学工具。随着质谱仪器和软件的改进,糖蛋白质组学的研究兴趣持续增长,因此,本综述旨在帮助研究人员获得必要的信息,选择最合适的富集策略,以最好地补充这些努力。