Quantitative chemical proteomics reveals mechanisms of action of clinical ABL kinase inhibitors
Quantitative chemical proteomics reveals mechanisms of action of clinical ABL kinase inhibitors
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DOI：
10.1038/nbt1328
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发表时间：
2007-09-01
影响因子：
46.9
通讯作者：
Drewes, Gerard
中科院分区：
文献类型：
--
作者：
Bantscheff, Marcus;Eberhard, Dirk;Drewes, Gerard
我们描述了一种化学蛋白质组学的方法来分析小分子与数百种内源性表达的蛋白激酶和嘌呤结合蛋白的相互作用。该亚蛋白质组被固定的非选择性激酶抑制剂（kinobeads）捕获，结合的蛋白质通过质谱法使用同量异位素标签进行相对和绝对定量（iTRAQ）平行定量。通过测量与亲和基质的竞争，我们评估了药物与其在细胞裂解物和细胞中的靶标的结合。通过绘制药物诱导的捕获蛋白质组磷酸化状态的变化，我们还分析了靶激酶下游的信号通路。药物伊马替尼（Gleevec），达沙替尼（Sprycel）和博舒替尼在K562细胞中的定量分析证实了已知的靶点，包括ABL和SRC家族激酶，并鉴定了受体酪氨酸激酶DDR1和氧化还原酶NQO2作为伊马替尼的新靶点。这些数据表明，我们的方法是药物发现的一个有价值的工具。
We describe a chemical proteomics approach to profile the interaction of small molecules with hundreds of endogenously expressed protein kinases and purine-binding proteins. This subproteome is captured by immobilized nonselective kinase inhibitors (kinobeads), and the bound proteins are quantified in parallel by mass spectrometry using isobaric tags for relative and absolute quantification (iTRAQ). By measuring the competition with the affinity matrix, we assess the binding of drugs to their targets in cell lysates and in cells. By mapping drug-induced changes in the phosphorylation state of the captured proteome, we also analyze signaling pathways downstream of target kinases. Quantitative profiling of the drugs imatinib (Gleevec), dasatinib (Sprycel) and bosutinib in K562 cells confirms known targets including ABL and SRC family kinases and identifies the receptor tyrosine kinase DDR1 and the oxidoreductase NQO2 as novel targets of imatinib. The data suggest that our approach is a valuable tool for drug discovery.