Isobaric labeling empowers proteome-wide expression measurements simultaneously across multiple samples. Here an expanded set of 16 isobaric reagents based on an isobutyl-proline immonium ion reporter structure (TMTpro) is presented. These reagents have similar characteristics to existing tandem mass tag reagents but with increased fragmentation efficiency and signal. In a proteome-scale example dataset, we compared eight common cell lines with and without Torin1 treatment with three replicates, quantifying more than 8,800 proteins (mean of 7.5 peptides per protein) per replicate with an analysis time of only 1.1 h per proteome. Finally, we modified the thermal stability assay to examine proteome-wide melting shifts after treatment with DMSO, 1 or 20 µM staurosporine with five replicates. This assay identified and dose-stratified staurosporine binding to 228 cellular kinases in just one, 18-h experiment. TMTpro reagents allow complex experimental designs-all with essentially no missing values across the 16 samples and no loss in quantitative integrity.

中文翻译：

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TMTpro试剂：一组同量异位标记质量标签，可同时对16个样品进行蛋白质组范围的测量。

等压标记可同时对多个样品进行蛋白质组范围内的表达测量。在此介绍了一组扩展的基于异丁基-脯氨酸铵离子报告结构（TMTpro）的16种同量异位试剂。这些试剂具有与现有串联质量标签试剂相似的特性，但具有更高的裂解效率和信号强度。在一个蛋白质组规模的示例数据集中，我们比较了使用和不使用Torin1处理的三种常见细胞系（三个重复），每个重复定量了8,800多种蛋白质（每个蛋白质7.5个肽的平均值），每个蛋白质组的分析时间仅为1.1小时。最后，我们修改了热稳定性测定法，以检查用DMSO，1或20 µM星形孢菌素处理5次重复后蛋白质组范围的融解。该试验仅在一个18小时的实验中就确定了星形孢菌素与228种细胞激酶的结合剂量分层。TMTpro试剂可进行复杂的实验设计-所有这一切在16个样品中基本没有缺失值，并且定量完整性也没有损失。