Novikova Svetlana E, Tikhonova Olga V, Kurbatov Leonid K, Farafonova Tatiana E, Vakhrushev Igor V, Zgoda Victor G
Institute of Biomedical Chemistry, Moscow, Russia.
Eur J Mass Spectrom (Chichester). 2017 Aug;23(4):202-208. doi: 10.1177/1469066717719848.
Targeted mass spectrometry represents a powerful tool for investigation of biological processes. The convenient approach of selected reaction monitoring using stable isotope-labeled peptide standard (SIS) is widely applied for protein quantification. Along with this method, high-resolution parallel reaction monitoring has been increasingly used for protein targeted analysis. Here we applied two targeted approaches (selected reaction monitoring with SIS and label-free parallel reaction monitoring) to investigate expression of 11 proteins during all-trans retinoic acid-induced differentiation of HL-60 cells. In our experiments, we have determined the proteins expression ratio at 3, 24, 48, and 96 h after all-trans retinoic acid treatment in comparison with 0 h, respectively. Expression profiles of four proteins (VAV1, PRAM1, LYN, and CEBPB) were highly correlated ( r > 0.75) and FGR expression was detected on proteome level starting from 24 h by both techniques. For prominent differences (fold change ≥ 2) label-free parallel reaction monitoring is not inferior to selected reaction monitoring with isotopically labeled peptide standards. Differentially expressed proteins, that have been determined in our study, can be considered as potential drug targets for acute myeloid leukemia (AML) treatment.
靶向质谱分析是研究生物过程的有力工具。使用稳定同位素标记肽标准品（SIS）的选择反应监测这种便捷方法被广泛应用于蛋白质定量分析。除了这种方法，高分辨率平行反应监测也越来越多地用于蛋白质靶向分析。在此，我们应用两种靶向方法（使用SIS的选择反应监测和无标记平行反应监测）来研究全反式维甲酸诱导HL-60细胞分化过程中11种蛋白质的表达情况。在我们的实验中，我们分别测定了全反式维甲酸处理后3、24、48和96小时与0小时相比的蛋白质表达比率。四种蛋白质（VAV1、PRAM1、LYN和CEBPB）的表达谱高度相关（r>0.75），并且两种技术均从24小时开始在蛋白质组水平检测到FGR表达。对于显著差异（倍数变化≥2），无标记平行反应监测并不逊色于使用同位素标记肽标准品的选择反应监测。我们研究中确定的差异表达蛋白质可被视为急性髓系白血病（AML）治疗的潜在药物靶点。
