Molecular Structure Analysis, German Cancer Research Center, Heidelberg, Germany.
Proteomics. 2010 Feb;10(4):634-49. doi: 10.1002/pmic.200900459.
The current status of de novo sequencing of peptides by MS/MS is reviewed with focus on collision cell MS/MS spectra. The relation between peptide structure and observed fragment ion series is discussed and the exhaustive extraction of sequence information from CID spectra of protonated peptide ions is described. The partial redundancy of the extracted sequence information and a high mass accuracy are recognized as key parameters for dependable de novo sequencing by MS. In addition, the benefits of special techniques enhancing the generation of long uninterrupted fragment ion series for de novo peptide sequencing are highlighted. Among these are terminal (18)O labeling, MS(n) of sodiated peptide ions, N-terminal derivatization, the use of special proteases, and time-delayed fragmentation. The emerging electron transfer dissociation technique and the recent progress of MALDI techniques for intact protein sequencing are covered. Finally, the integration of bioinformatic tools into peptide de novo sequencing is demonstrated.
本文综述了通过 MS/MS 对肽进行从头测序的现状，重点讨论了碰撞池 MS/MS 谱。讨论了肽结构与观察到的片段离子系列之间的关系，并描述了从质子化肽离子的 CID 谱中彻底提取序列信息。提取的序列信息的部分冗余性和高质量精度被认为是通过 MS 进行可靠从头测序的关键参数。此外，还强调了增强用于从头测序的肽的长连续片段离子系列生成的特殊技术的优势。其中包括末端（18）O 标记、加钠肽离子的 MS(n)、N 末端衍生化、特殊蛋白酶的使用以及延迟碎裂。涵盖了新兴的电子转移解离技术以及 MALDI 技术在完整蛋白质测序方面的最新进展。最后，展示了将生物信息学工具整合到肽从头测序中的方法。
