Tracking cancer drugs in living cells by thermal profiling of the proteome
Tracking cancer drugs in living cells by thermal profiling of the proteome
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DOI：
10.1126/science.1255784
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发表时间：
2014-10-03
期刊：
影响因子：
56.9
通讯作者：
Drewes, Gerard
中科院分区：
文献类型：
--
作者：
Savitski, Mikhail M.;Reinhard, Friedrich B. M.;Drewes, Gerard
蛋白质的热稳定性可用于评估配体在活细胞中的结合。我们通过质谱法测定人体细胞中7000多种蛋白质的热谱，概括了这一概念。监测小分子配体对激酶抑制剂staurosporine的50多个靶点的影响。我们发现血红素生物合成酶铁螯合酶是激酶抑制剂的靶标，并认为其抑制导致了vemurafenib和alectinib所观察到的光毒性。在药物治疗的下游效应物中也观察到热位移。在活细胞中，达沙替尼诱导BCR-ABL通路蛋白的转移，包括CRK/CRKL。热蛋白质组分析提供了一种无偏倚的药物靶标参与测量方法，并促进了药物疗效和毒性标记物的鉴定。
The thermal stability of proteins can be used to assess ligand binding in living cells. We have generalized this concept by determining the thermal profiles of more than 7000 proteins in human cells by means of mass spectrometry. Monitoring the effects of small-molecule ligands on the profiles delineated more than 50 targets for the kinase inhibitor staurosporine. We identified the heme biosynthesis enzyme ferrochelatase as a target of kinase inhibitors and suggest that its inhibition causes the phototoxicity observed with vemurafenib and alectinib. Thermal shifts were also observed for downstream effectors of drug treatment. In live cells, dasatinib induced shifts in BCR-ABL pathway proteins, including CRK/CRKL. Thermal proteome profiling provides an unbiasedmeasure of drug-target engagement and facilitates identification of markers for drug efficacy and toxicity.