Increased N,N-Dimethyl Leucine Isobaric Tag Multiplexing by a Combined Precursor Isotopic Labeling and Isobaric Tagging Approach,Analytical Chemistry

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Increased N,N-Dimethyl Leucine Isobaric Tag Multiplexing by a Combined Precursor Isotopic Labeling and Isobaric Tagging Approach
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-08-10 00:00:00 , DOI: 10.1021/acs.analchem.8b01301
Dustin C. Frost ₁ , Clayton J. Rust ₂ , Renã A. S. Robinson ₃ , Lingjun Li _{1,

2}

Affiliation

Multiplex isobaric tags have become valuable tools for high-throughput quantitative analysis of complex biological samples in discovery-based proteomics studies. Hybrid labeling strategies that pair stable isotope mass difference labeling with multiplex isobaric tag-based quantification further facilitate these studies by greatly increasing multiplexing capability. In this work, we present a cost-effective chemical labeling approach that couples duplex stable isotope dimethyl labeling with our custom 12-plex N,N-dimethyl leucine (DiLeu) isobaric tags in a combined precursor isotopic labeling and isobaric tagging (cPILOT) strategy that is compatible with a wide variety of biological samples and permits 24-plex quantification in a single LC-MS/MS experiment. We demonstrate the utility of the DiLeu cPILOT approach by labeling yeast digests and performing proof-of-principle quantification experiments on the Orbitrap Fusion Lumos.

中文翻译：

前体同位素标记和等压标记法相结合的方法增加了N，N-二甲基亮氨酸等压标记的多路复用

在基于发现的蛋白质组学研究中，多重等压标记物已成为对复杂生物样品进行高通量定量分析的有价值的工具。将稳定同位素质量差异标记与基于多重等压标记的定量配对的混合标记策略通过大大提高多重能力进一步促进了这些研究。在这项工作中，我们提出了一种经济有效的化学标记方法，该方法将双链稳定同位素二甲基标记与我们定制的12重N，N偶联-前体同位素标记和等压标记（cPILOT）组合策略中的-二甲基亮氨酸（DiLeu）等压标记，可与多种生物样品兼容，并允许在单个LC-MS / MS实验中进行24多重定量。我们通过标记酵母消化物并在Orbitrap Fusion Lumos上进行原理证明量化实验来证明DiLeu cPILOT方法的实用性。

更新日期：2018-08-10

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