Wuhrer Manfred, Catalina M Isabel, Deelder André M, Hokke Cornelis H
Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Apr 15;849(1-2):115-28. doi: 10.1016/j.jchromb.2006.09.041. Epub 2006 Oct 17.
Next to the identification of proteins and the determination of their expression levels, the analysis of post-translational modifications (PTM) is becoming an increasingly important aspect in proteomics. Here, we review mass spectrometric (MS) techniques for the study of protein glycosylation at the glycopeptide level. Enrichment and separation techniques for glycoproteins and glycopeptides from complex (glyco-)protein mixtures and digests are summarized. Various tandem MS (MS/MS) techniques for the analysis of glycopeptides are described and compared with respect to the information they provide on peptide sequence, glycan attachment site and glycan structure. Approaches using electrospray ionization and matrix-assisted laser desorption/ionization (MALDI) of glycopeptides are presented and the following fragmentation techniques in glycopeptide analysis are compared: collision-induced fragmentation on different types of instruments, metastable fragmentation after MALDI ionization, infrared multi-photon dissociation, electron-capture dissociation and electron-transfer dissociation. This review discusses the potential and limitations of tandem mass spectrometry of glycopeptides as a tool in structural glycoproteomics.
除了蛋白质鉴定及其表达水平的测定，翻译后修饰（PTM）分析正成为蛋白质组学中一个日益重要的方面。在此，我们综述了用于在糖肽水平研究蛋白质糖基化的质谱（MS）技术。总结了从复杂（糖）蛋白混合物和消化物中富集和分离糖蛋白及糖肽的技术。描述了用于分析糖肽的各种串联质谱（MS/MS）技术，并就它们在肽序列、聚糖连接位点和聚糖结构方面提供的信息进行了比较。介绍了使用电喷雾电离和糖肽的基质辅助激光解吸/电离（MALDI）的方法，并比较了糖肽分析中的以下裂解技术：不同类型仪器上的碰撞诱导裂解、MALDI电离后的亚稳裂解、红外多光子解离、电子捕获解离和电子转移解离。本综述讨论了糖肽串联质谱作为结构糖蛋白质组学工具的潜力和局限性。
