De novo sequencing of peptides by MS/MS
2009; Wiley; Volume: 10; Issue: 4 Linguagem: Inglês
10.1002/pmic.200900459
ISSN1615-9861
AutoresJoerg Seidler, Nico Zinn, Martin E. Boehm, Wolf D. Lehmann,
Tópico(s)Molecular Biology Techniques and Applications
ResumoThe 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.
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