Medzihradszky Katalin F
Department of Pharmaceutical Chemistry, University of California, San Francisco, USA.
Methods Enzymol. 2005;402:209-44. doi: 10.1016/S0076-6879(05)02007-0.
In mass spectrometry (MS)-based protein studies, peptide fragmentation analysis (i.e., MS/MS experiments such as matrix-assisted laser desorption ionization [MALDI]-post-source decay [PSD] analysis, collision-induced dissociation [CID] of electrospray- and MALDI-generated ions, and electron-capture and electron-transfer dissociation analysis of multiply charged ions) provide sequence information and, thus, can be used for (i) de novo sequencing, (ii) protein identification, and (iii) posttranslational or other covalent modification site assignments. This chapter offers a qualitative overview on which kind of peptide fragments are formed under different MS/MS conditions. High-quality PSD and CID spectra provide illustrations of de novo sequencing and protein identification. The MS/MS behavior of some common posttranslational modifications such as acetylation, trimethylation, phosphorylation, sulfation, and O-glycosylation is also discussed.
在基于质谱(MS)的蛋白质研究中,肽段碎裂分析(即MS/MS实验,如基质辅助激光解吸电离[MALDI]源后衰变[PSD]分析、电喷雾和MALDI产生离子的碰撞诱导解离[CID],以及多电荷离子的电子捕获和电子转移解离分析)可提供序列信息,因此可用于:(i) 从头测序;(ii) 蛋白质鉴定;(iii) 翻译后或其他共价修饰位点的确定。本章对在不同MS/MS条件下形成的肽段碎片类型进行了定性概述。高质量的PSD和CID谱图展示了从头测序和蛋白质鉴定过程。还讨论了一些常见翻译后修饰(如乙酰化、三甲基化、磷酸化、硫酸化和O-糖基化)的MS/MS行为。