Targeted Quantitative Proteomics for the Analysis of 14 UGT1As and-2Bs in Human Liver Using NanoUPLC-MS/MS with Selected Reaction Monitoring
Targeted Quantitative Proteomics for the Analysis of 14 UGT1As and-2Bs in Human Liver Using NanoUPLC-MS/MS with Selected Reaction Monitoring
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DOI：
10.1021/pr4004213
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发表时间：
2013-10-01
影响因子：
4.4
通讯作者：
Smith, Philip C.
中科院分区：
文献类型：
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作者：
Fallon, John K.;Neubert, Hendrik;Smith, Philip C.
使用重同位素稀释技术的靶向定量蛋白质组学越来越多地用于定量生物基质中的蛋白质，包括 UGT 酶。在这里，我们提出了一种使用 nanoLC-MS/MS 和多反应监测 (MRM) 的多重方法来定量肝脏基质中的 14 个 UGT1A 和 UGT2B。在可行的情况下，我们对每种蛋白质使用两种或多种蛋白型肽，仅用一种蛋白型肽对四种蛋白质进行定量。我们应用该方法分析 60 个人类肝微粒体 (HLM) 库和匹配的 S9 样本。在肝脏中可检测到10种UGT亚型，每种亚型的表达与文献报道的mRNA表达一致。 UGT2B17 的不寻常之处在于，大约 30% 的肝微粒体没有或很少有（
Targeted quantitative proteomics using heavy isotope dilution techniques is increasingly being utilized to quantify proteins, including UGT enzymes, in biological matrices. Here we present a multiplexed method using nanoLC-MS/MS and multiple reaction monitoring (MRM) to quantify 14 UGT1As and UGT2Bs in liver matrices. Where feasible, we employ two or more proteotypic peptides per protein, with only four proteins quantified with only one proteotypic peptide. We apply the method to analysis of a library of 60 human liver microsome (HLM) and matching S9 samples. Ten of the UGT isoforms could be detected in liver, and the expression of each was consistent with mRNA expression reported in the literature. UGT2B17 was unusual in that similar to 30% of liver microsomes had no or little (