Jang-Lee Jihye, North Simon J, Sutton-Smith Mark, Goldberg David, Panico Maria, Morris Howard, Haslam Stuart, Dell Anne
Department of Molecular Biosciences, Imperial College London, UK.
Methods Enzymol. 2006;415:59-86. doi: 10.1016/S0076-6879(06)15005-3.
Over the past decade, rapid, high-sensitivity mass spectrometric strat-egies have been developed and optimized for screening for the types of N- and O-glycans present in a diverse range of biological material, including secretions, cell lines, tissues, and organs. These glycomic strategies are based on matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass fingerprinting of permethylated derivatives, combined with electrospray (ES) or MALDI tandem mass spectrometry (MS/MS) sequencing and gas chromatography (GC)-MS linkage analysis, complemented by chemical and enzymatic degradations. Protocols for these methods are described in the first part of this chapter. Glycomic experiments yield large volumes of MS data, and interpretation of the resulting spectra remains a time-consuming bottleneck in the process. In the second part of this chapter, we describe the use and operation of a mass spectral viewer program capable of displaying and automatically labeling spectra arising from MALDI fingerprinting of N-glycans.
在过去十年中,已开发并优化了快速、高灵敏度的质谱策略,用于筛查多种生物材料(包括分泌物、细胞系、组织和器官)中存在的N-聚糖和O-聚糖类型。这些糖组学策略基于对全甲基化衍生物的基质辅助激光解吸/电离(MALDI)飞行时间质谱指纹图谱,结合电喷雾(ES)或MALDI串联质谱(MS/MS)测序以及气相色谱(GC)-MS关联分析,并辅以化学和酶促降解。本章第一部分介绍了这些方法的实验方案。糖组学实验会产生大量的质谱数据,而对所得光谱的解读仍然是该过程中耗时的瓶颈。在本章第二部分,我们描述了一种质谱查看器程序的使用和操作,该程序能够显示并自动标记由N-聚糖的MALDI指纹图谱产生的光谱。