Department of Chemistry, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.
Proteomics. 2011 Feb;11(4):631-43. doi: 10.1002/pmic.201000517. Epub 2011 Jan 17.
The oligosaccharides attached to proteins or lipids are among the most challenging analytical tasks due to their complexity and variety. Knowing the genes and enzymes responsible for their biosynthesis, a large but not unlimited number of different structures and isomers of such glycans can be imagined. Understanding of the biological role of structural variations requires the ability to unambiguously determine the identity and quantity of all glycan species. Here, we examine, which analytical strategies - with a certain high-throughput potential - may come near this ideal. After an expose of the relevant techniques, we try to depict how analytical raw data are translated into structural assignments using retention times, mass and fragment spectra. A method's ability to discriminate between the many conceivable isomeric structures together with the time, effort and sample amount needed for that purpose is suggested as a criterion for the comparative assessment of approaches and their evolutionary stages.
由于其复杂性和多样性,附着在蛋白质或脂质上的寡糖是最具挑战性的分析任务之一。了解负责其生物合成的基因和酶,可以想象出大量但不是无限数量的不同结构和此类聚糖的异构体。对结构变异的生物学作用的理解需要能够明确确定所有聚糖种类的身份和数量。在这里,我们研究了哪些分析策略 - 具有一定的高通量潜力 - 可能接近这一理想。在介绍了相关技术之后,我们尝试描述如何使用保留时间、质量和碎片谱将分析原始数据转换为结构分配。该方法区分许多可想象的异构结构的能力以及为此目的所需的时间、精力和样品量被建议作为比较评估方法及其进化阶段的标准。