Carboxy group derivatization for enhanced electron-transfer dissociation mass spectrometric analysis of phosphopeptides.
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.
Zhang
Lijuan
L
Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, PR China.
Xu
Yawei
Y
Lu
Haojie
H
Yang
Pengyuan
P
D002065:Buspirone / Q000031:analogs & derivatives / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010748:Phosphopeptides / Q000737:chemistry*
D010748:Phosphopeptides; C037921:1-(2-pyrimidinyl)piperazine; D002065:Buspirone