Comparative Evaluation of Two Isobaric Labeling Tags, DiART and iTRAQ
Comparative Evaluation of Two Isobaric Labeling Tags, DiART and iTRAQ
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DOI：
10.1021/ac203467q
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发表时间：
2012-03-20
影响因子：
7.4
通讯作者：
Liu, Siqi
中科院分区：
文献类型：
--
作者：
Chen, Zhen;Wang, Quanhui;Liu, Siqi
等压标签在基础和转化研究中都有广泛的应用，如广泛使用的相对和绝对定量等压标签（iTRAQ）。然而，最近大规模定量蛋白质组学项目的结果表明，iTRAQ的蛋白质定量在复杂的生物样品中往往存在偏差。在这里，我们报道了另一种等压标签的应用，氘等压胺反应标签（DiART），用于定量热厌氧细菌tengcongensis （T. tengcongensis）的蛋白质组，这是中国首次发现的一种嗜热细菌。我们从几个不同的方面比较了DiART和iTRAQ的性能，包括它们的断裂机制、鉴定的蛋白质数量和定量的准确性。我们的研究结果表明，与iTRAQ相比，DiART产生的报告离子明显更强，这没有减少可识别肽的数量，但提高了量化的信噪比（S/N）。值得注意的是，我们发现，在相同的色谱和质谱（MS）条件下，DiART比iTRAQ表现出更少的报告离子比压缩，这可能是由于DiART标记产生的报告离子更多，强度更高。综上所述，我们证明DiART是iTRAQ的一个有价值的替代品，具有增强的定量蛋白质组学性能。
Isobaric tags have broad applications in both basic and translational research, as demonstrated by the widely used isobaric tag for relative and absolute quantitation (iTRAQ). Recent results from large-scale quantitative proteomics projects, however, indicate that protein quantification by iTRAQ is often biased in complex biological samples. Here, we report the application of another isobaric tag, deuterium isobaric amine reactive tag (DiART), for quantifying the proteome of Thermoanaerobacter tengcongensis (T. tengcongensis), a thermophilic bacterium first discovered in China. We compared the performance of DiART with iTRAQ from several different aspects, including their fragmentation mechanisms, the number of identified proteins, and the accuracy of quantification. Our results revealed that, as compared with iTRAQ DiART yielded significantly stronger reporter ions, which did not reduce the number of identifiable peptides, but improved the signal-to-noise ratio (S/N) for quantification. Remarkably, we found that, under identical chromatography and mass spectrometry (MS) conditions, DiART exhibited less reporter ions ratio compression than iTRAQ probably due to more reporter ions with higher intensities produced by DiART labeling. Taken together, we demonstrate that DiART is a valuable alternative of iTRAQ with enhanced performance for quantitative proteomics.