A novel triplex isobaric termini labeling quantitative approach for simultaneously supplying three quantitative sources
A novel triplex isobaric termini labeling quantitative approach for simultaneously supplying three quantitative sources
复制标题
一种新颖的三重同量异位末端标记定量方法，用于同时提供三个定量源。
DOI：
10.1016/j.aca.2017.11.004
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发表时间：
2018-02-25
影响因子：
6.2
通讯作者：
Lu, Haojie
中科院分区：
文献类型：
--
作者：
Jiang, Hucong;Yin, Hongrui;Lu, Haojie
Benefiting from high sensitivity and great ability to measure multiple samples simultaneously, isobaric tandem Mass spectrometry (MS2) quantification has been widely applied for protein biomarker screening. Here, a newly developed isobaric MS2 quantification method named triplex quantification by isobaric termini labeling (Triplex-QITL) was established. This method enables the accurate comparison of various fragment ions (reporter ions, amino acid fragments and N-/C-terminal fragments) based quantification to be operated in a single run. To our knowledge, this is the first time that this kind of comparison is achieved. In Triplex-QITL, proteins were first digested with Lys-C to produce peptides with lysine (K) at the C-termini, then dimethylation reagents and mTRAQ reagents were used to label the N-termini and C-termini of the peptides respectively. N- and C-terminal fragment ion pairs, reporter ions from mTRAQ (113,117,121) and a(1) ion pairs were simultaneously generated in MS2 spectra. In simple sample experiment, not much difference in using various fragment ions for quantification was observed. When analyzing SW480 cell lysate, comparing with a1 ions, about two times of reproducible quantification results were achieved by reporter ions and N- and C-terminal ions. Meanwhile the measured quantification results were much closer to the expected results even in large ratios (1:10:10) using N- and C-terminal ions. Finally, Triplex-QITL was successfully applied to profile metastatic differences of three hepatocellular carcinoma (HCC) cell lines. In all, Triplex-QITL shows a promising future in quantitative proteomics. (c) 2017 Elsevier B.V. All rights reserved.
