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Evaluation of the accuracy of protein quantification using isotope TMPP‐labeled peptides
Author(s) -
Shen Hongyan,
An Mingrui,
Zou Xiao,
Zhao Xuyang,
Wang Qingsong,
Xing Guowen,
Ji Jianguo
Publication year - 2015
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.201400495
Subject(s) - quantitative proteomics , derivatization , stable isotope labeling by amino acids in cell culture , chemistry , peptide , chromatography , proteomics , phosphonium , isobaric labeling , isotope , bromide , isotopic labeling , label free quantification , mass spectrometry , biochemistry , organic chemistry , physics , quantum mechanics , gene
N ‐succinimidyloxycarbonylmethyl tris (2,4,6‐trimethoxyphenyl) phosphonium bromide (TMPP‐Ac‐OSu) reacts rapidly, mildly, and specifically with the N‐terminals of proteins and peptides. Thus, it can be developed as an ideal isotope‐coded tag to be used in quantitative proteomics. Here, we present a strategy for light and heavy TMPP‐based quantitative proteomic analysis, in which peptides in a mixture can be quantified using an on‐tip TMPP derivatization approach. To demonstrate the accuracy of this strategy, light and heavy TMPP‐labeled peptides were combined at different ratios and subsequently analyzed by LC‐MS/MS. The MS spectra and scatter plots show that peptide and protein ratios were both consistent with the mixed ratios. We observed a linear correlation between protein ratios and the predicted ratios. In comparison with SILAC method, the TMPP labeling method produced similarly accurate quantitative results with low CVs. In conclusion, our results suggest that this isotope‐coded TMPP method achieved accurate quantification and compatibility with IEF‐based separation. With the inherent advantages of TMPP derivatization, we believe that it holds great promise for future applications in quantitative proteomics analysis.