Deuterium-Free, Three-Plexed Peptide Diethylation for Highly Accurate Quantitative Proteomics.,Journal of Proteome Research

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Deuterium-Free, Three-Plexed Peptide Diethylation for Highly Accurate Quantitative Proteomics.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2019-01-29 , DOI: 10.1021/acs.jproteome.8b00775
Jaehun Jung ₁ , Kyowon Jeong _{2,

3} , Yeon Choi _{2,

3} , Sun Ah Kim _{2,

3} , Hyunjoon Kim _{2,

3} , Joon Won Lee ₁ , V Narry Kim _{2,

3} , Kwang Pyo Kim ₁ , Jong-Seo Kim _{2,

3}

Affiliation

The deuterium, a frequently used stable isotope in isotopic labeling for quantitative proteomics, could deteriorate the accuracy and precision of proteome quantification owing to the retention time shift of deuterated peptides from the hydrogenated counterpart. We introduce a novel three-plexed peptide "diethylation" using only 13C isotopologues of acetaldehyde and demonstrate that the accuracy and precision of our method in proteome quantification are significantly superior to the conventional deuterium-based dimethylation labeling in both a single-shot and multidimensional LC-MS/MS analysis of the HeLa proteome. Furthermore, in time-resolved profiling of Xenopus laevis early embryogenesis, our 3-plexed diethylation outperformed isobaric labeling approaches in terms of the quantification accuracy or the number of protein identifications, generating more than two times more differentially expressed proteins. Our cost-effective and highly accurate 3-plexed diethylation method could contribute to various types of quantitative proteomics applications in which three of multiplexity would be sufficient.

中文翻译：

不含氘的三重肽二乙基化，用于高精度定量蛋白质组学。

氘是在同位素标记中用于定量蛋白质组学的常用稳定同位素，由于氘代肽段与氢化对应物之间的保留时间发生偏移，氘可能会降低蛋白质组定量的准确性和准确性。我们仅使用乙醛的13 C同位素，介绍了一种新颖的三重肽“二乙基化”，并证明了我们的蛋白质组定量方法的准确性和精密度在单次和多维LC中均显着优于传统的基于氘的二甲基化标记HeLa蛋白质组的-MS / MS分析。此外，在Xenopus laevis早期胚胎发生的时间分辨谱分析中，就定量准确度或蛋白质鉴定数量而言，我们的3联二乙基化性能优于同量异位标记方法，产生两倍以上的差异表达蛋白质。我们具有成本效益且高度准确的3-plex diethylation方法可为多种类型的定量蛋白质组学应用做出贡献，其中三种复用就足够了。

更新日期：2019-02-07

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