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Updated: Jun 21, 2026

Covalent Labeling with Diethylpyrocarbonate for Studying Protein Higher-Order Structure by Mass Spectrometry
Published on: June 15, 2021
Carboxy group derivatization for enhanced electron-transfer dissociation mass spectrometric analysis of
Lijuan Zhang1, Yawei Xu, Haojie Lu
1Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, PR China.
Abstract:
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.
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