Efficient and clean charge derivatization of peptides for analysis by mass spectrometry

摘要: Charge derivatization with succinimidyloxycarbonylmethyl tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP-Ac-OSu) has great potential in several aspects of proteomics, such as peptide de novo sequencing, PTM analysis, etc. However, the excess reagent and its side products greatly limited scope use. Here, we report an improved method to perform charge peptides by TMPP-Ac-OSu without interference from corresponding products. Briefly, protein was first separated on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or coagulated gel. The in-gel then incubated a high concentration reagent, followed extensive washing. Afterwards, digested trypsin according routine protocol. mainly resultant were attached one positive tag N-termini Lys-epsilon-NH(2). process been successfully applied 2-DE resolved spots. Compared native proteins, derivatized counterparts have higher rates PMF identification more straightforward tandem mass spectra. This promising should pave way for practical use proteomics.