Differential Protein Expression Analysis Using Stable Isotope Labeling and PQD Linear Ion Trap MS Technology
Differential Protein Expression Analysis Using Stable Isotope Labeling and PQD Linear Ion Trap MS Technology
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DOI：
10.1016/j.jasms.2009.02.029
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发表时间：
2009-07-01
影响因子：
3.2
通讯作者：
Lazar, Iulia M.
中科院分区：
文献类型：
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作者：
Armenta, Jenny M.;Hoeschele, Ina;Lazar, Iulia M.
建立了基于相对和绝对定量同位素标记(ITRAQ)的反相高效液相(RPLC)-串联质谱(MS/MS)方法，用于复杂细胞提取液中蛋白质的差异表达谱分析。以雌激素阳性的MCF-7细胞株为模型，在17β-雌二醇(E_2)和三苯氧胺()的共同作用下培养。采用脉冲Q离解(PQD)检测的线性陷阱四极(LTQ)仪进行MS分析。通过优化实验，最大限度地提高了iTRAQ的标记效率和定量蛋白质的数量。选择MS数据过滤标准以导致假阳性识别率为
An isotope tags for relative and absolute quantitation (iTRAQ)-based reversed-phase liquid chromatography (RPLC)-tandem mass spectrometry (MS/MS) method was developed for differential protein expression profiling in complex cellular extracts. The estrogen positive MCF-7 cell line, cultured in the presence of 17 beta-estradiol (E2) and tamoxifen (Tam), was used as a model system. MS analysis was performed with a linear trap quadrupole (LTQ) instrument operated by using pulsed Q dissociation (PQD) detection. Optimization experiments were conducted to maximize the iTRAQ labeling efficiency and the number of quantified proteins. MS data filtering criteria were chosen to result in a false positive identification rate of