A Pragmatic Guide to Enrichment Strategies for Mass Spectrometry-Based Glycoproteomics.
A Pragmatic Guide to Enrichment Strategies for Mass Spectrometry-Based Glycoproteomics.
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DOI：
10.1074/mcp.r120.002277
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Pitteri SJ
中科院分区：
文献类型：
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作者：
Riley NM;Bertozzi CR;Pitteri SJ
糖基化是一种普遍存在的异质性修饰，在分子生物学中具有广泛的意义。这种异质性排除了对所有聚糖类别普遍有益的富集策略。因此，富集策略的选择对实验结果有着深远的影响。在这里，我们回顾了现代质谱为基础的糖蛋白质组学实验中使用的常见富集策略，包括凝集素和其他亲和色谱，亲水相互作用色谱及其衍生物，多孔石墨碳，可逆和不可逆的化学偶联策略，和化学生物学工具，往往利用生物正交手柄。随着质谱仪器和软件的改进，对糖蛋白组学的兴趣不断增加，因此本文旨在帮助研究人员获得必要的信息，以选择最佳的富集策略来补充这些努力。糖基化是复杂的，往往需要富集分析前审查常见的富集策略，质谱为基础的糖蛋白质组学富集方法有实际的考虑和实验的影响适当的富集策略将补充质谱的发展基于质谱的糖蛋白质组学分析的兴趣正在增加，因为最近的进展仪器和数据分析工具。这些研究可以提供广泛的聚糖类和生物系统的丰富信息。然而，许多研究需要在分析之前选择糖基化物质的富集策略，以获得最大量的分析信息。在这里，常见的富集策略的优点和缺点进行了审查，并讨论了各种方法的实际考虑。
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. Glycosylation is complex and often requires enrichment prior to analysis Review of common enrichment strategies for mass spectrometry–based glycoproteomics Enrichment methods have practical considerations and experimental implications Appropriate enrichment strategies will complement developments in mass spectrometry Interest in mass spectrometry–based glycoproteomics analysis is increasing because of recent advances in instrumentation and data analysis tools. Such studies can provide a wealth of information across a wide spectrum of glycan classes and biological systems. However, many studies require the choice of an enrichment strategy for glycosylated species prior to analysis to obtain the maximum amount of analytical information. Here, common enrichment strategies are reviewed with strengths and weaknesses, and the practical considerations for various methods are discussed.