Chaperones as thermodynamic sensors of drug-target interactions reveal kinase inhibitor specificities in living cells.
Chaperones as thermodynamic sensors of drug-target interactions reveal kinase inhibitor specificities in living cells.
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DOI：
10.1038/nbt.2620
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发表时间：
2013-07
影响因子：
46.9
通讯作者：
中科院分区：
文献类型：
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HSP90分子伴侣与其客户激酶之间的相互作用对激酶的构象状态敏感，并且小分子对激酶折叠的稳定性强烈降低分子伴侣相互作用。在这里，我们利用这一观察和分析小分子结合激酶在活细胞中，使用分子伴侣作为“热力学传感器”。该方法允许在激酶的天然细胞环境中以高通量测定ATP竞争性和变构抑制剂的靶特异性。我们分析了30种不同激酶抑制剂对> 300种激酶的靶特异性。为了证明该测定的价值，我们将ETV6-NTRK3确定为FDA批准的药物克唑替尼（Xalkori）的靶标。克唑替尼以纳摩尔效力抑制ETV6-NTRK3依赖性肿瘤细胞的增殖，并诱导小鼠中已建立的肿瘤异种移植物消退。最后，我们通过分析HSP 70/类固醇激素受体和CDC37/激酶的相互作用表明，我们的方法适用于其他分子伴侣和靶类，这表明分子伴侣相互作用将在体内检测药物-靶相互作用中具有广泛的应用。
The interaction between the HSP90 chaperone and its client kinases is sensitive to the conformational status of the kinase, and stabilization of the kinase fold by small molecules strongly decreases chaperone interaction. Here we exploit this observation and assay small-molecule binding to kinases in living cells, using chaperones as 'thermodynamic sensors'. The method allows determination of target specificities of both ATP-competitive and allosteric inhibitors in the kinases' native cellular context in high throughput. We profile target specificities of 30 diverse kinase inhibitors against >300 kinases. Demonstrating the value of the assay, we identify ETV6-NTRK3 as a target of the FDA-approved drug crizotinib (Xalkori). Crizotinib inhibits proliferation of ETV6-NTRK3-dependent tumor cells with nanomolar potency and induces the regression of established tumor xenografts in mice. Finally, we show that our approach is applicable to other chaperone and target classes by assaying HSP70/steroid hormone receptor and CDC37/kinase interactions, suggesting that chaperone interactions will have broad application in detecting drug-target interactionsin vivo.
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影响因子：
5.1
作者：
Atanassov II;Atanassov II;Etchells JP;Turner SR
通讯作者：
Turner SR
影响因子：
46.9
作者：
通讯作者：
--
影响因子：
64.5
作者：
Azam, M;Latek, RR;Daley, GQ
通讯作者：
Daley, GQ
影响因子：
5.7
作者：
Gozgit JM;Wong MJ;Wardwell S;Tyner JW;Loriaux MM;Mohemmad QK;Narasimhan NI;Shakespeare WC;Wang F;Druker BJ;Clackson T;Rivera VM
通讯作者：
Rivera VM
DOI：
10.1073/pnas.0708800104
发表时间：
2007-12-18
影响因子：
11.1
作者：
Fedorov, Oleg;Marsden, Brian;Knapp, Stefan
通讯作者：
Knapp, Stefan