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Carboxy group derivatization for enhanced electron‐transfer dissociation mass spectrometric analysis of phosphopeptides
Author(s) -
Zhang Lijuan,
Xu Yawei,
Lu Haojie,
Yang Pengyuan
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800963
Subject(s) - derivatization , phosphopeptide , chemistry , electron transfer dissociation , chromatography , dissociation (chemistry) , combinatorial chemistry , mass spectrometry , tandem mass spectrometry , organic chemistry , peptide , biochemistry
A novel strategy based on carboxy group derivatization is presented for specific characterization of phosphopeptides. By tagging the carboxy group with 1‐(2‐pyrimidyl) piperazine (PP), the ion charge states of phosphopeptides can be largely enhanced, showing great advantages for sequencing phosphorylated peptides with electron‐transfer dissociation MS. Besides, after PP‐derivatization, most non‐specific bindings can be avoided by eliminating the interaction between the carboxy group and TiO 2 , greatly improving the specificity of TiO 2 ‐based phosphopeptide enrichment strategy. Moreover, being tagged with a hydrophobic group, the retention time of phosphopeptides in RPLC can be prolonged, overcoming the difficulty of separating phosphopeptides in RPLC‐based approach. Together with several other advantages, such as ease of handling, rapid reaction time, broad applicability and good reproducibility, this PP‐derivatization method is promising for high‐throughput phosphoproteome research.