Gatekeeper mutations activate FGF receptor tyrosine kinases by destabilizing the autoinhibited state.
Gatekeeper mutations activate FGF receptor tyrosine kinases by destabilizing the autoinhibited state.
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DOI：
10.1073/pnas.2213090120
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发表时间：
2023-02-21
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
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作者：
抑制受体酪氨酸激酶激酶结构域的抗癌药物阻断了细胞信号传导和生长所必需的下游磷酸化。然而，成功治疗癌症的一个主要障碍是在位于激酶铰区（位于N-叶和c -叶之间）的残基上出现耐药性突变，该残基为肿瘤生长提供了适应性优势。在这项工作中，我们使用激酶分析、核磁共振光谱和MD模拟来确定耐药看门人突变引起的激酶激活机制。我们发现这些突变通过削弱疏水脊和调节保守DFG基序的构象来破坏激酶的自抑制构象，从而得出结论，在活跃状态的人群中，守门人突变增加。许多类型的人类癌症正在使用靶向受体酪氨酸激酶的激酶结构域的小分子atp竞争性抑制剂进行治疗。尽管最初成功缓解，但长期治疗几乎不可避免地导致在看门人残基上出现耐药性突变，阻碍了抑制剂进入atp结合间隙后部的疏水口袋。除了降低药物疗效外，看门人突变还会提高酪氨酸激酶结构域的内在活性，从而导致更具侵袭性的癌症类型。然而，看门人突变的功能获得机制尚不清楚。在这里，我们使用激酶活性测定、核磁共振光谱、生物信息学分析和MD模拟来表征成纤维细胞生长因子受体（FGFR）酪氨酸激酶具有两种不同的看门人突变。我们的数据表明，看门人突变破坏了局部DFG基序和激酶整体的自抑制构象，表明它们通过促进激酶填充活性状态的能力来赋予功能获得。
Anticancer drugs inhibiting the kinase domain of receptor tyrosine kinases block downstream phosphorylation necessary for cellular signaling and growth. However, a major impediment to successful cancer therapy is the emergence of drug resistance mutations at a residue situated at the kinase hinge region between the N- and C-lobes, which provides fitness advantages for tumor growth. In this work, we used kinase assays, NMR spectroscopy, and MD simulations to determine the mechanism of kinase activation from drug-resistant gatekeeper mutations. We found that these mutations destabilize the autoinhibited conformation of the kinase by weakening the hydrophobic spine and modulating the conformation of the conserved DFG motif, thereby leading to the conclusion that gatekeeper mutations increase in the population of the active state. Many types of human cancers are being treated with small molecule ATP-competitive inhibitors targeting the kinase domain of receptor tyrosine kinases. Despite initial successful remission, long-term treatment almost inevitably leads to the emergence of drug resistance mutations at the gatekeeper residue hindering the access of the inhibitor to a hydrophobic pocket at the back of the ATP-binding cleft. In addition to reducing drug efficacy, gatekeeper mutations elevate the intrinsic activity of the tyrosine kinase domain leading to more aggressive types of cancer. However, the mechanism of gain-of-function by gatekeeper mutations is poorly understood. Here, we characterized fibroblast growth factor receptor (FGFR) tyrosine kinases harboring two distinct gatekeeper mutations using kinase activity assays, NMR spectroscopy, bioinformatic analyses, and MD simulations. Our data show that gatekeeper mutations destabilize the autoinhibitory conformation of the DFG motif locally and of the kinase globally, suggesting they impart gain-of-function by facilitating the kinase's ability to populate the active state.
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影响因子：
2.2
作者：
CARVER, JP;RICHARDS, RE
通讯作者：
RICHARDS, RE
影响因子：
16
作者：
Chen, Catherine;Ha, Byung Hak;Thevenin, Anastasia F.;Lou, Hua Jane;Zhang, Rong;Yip, Kevin Y.;Peterson, Jeffrey R.;Gerstein, Mark;Kim, Philip M.;Filippakopoulos, Panagis;Knapp, Stefan;Boggon, Titus J.;Turk, Benjamin E.
通讯作者：
Turk, Benjamin E.
影响因子：
5.3
作者：
Hu, Jiancheng;Ahuja, Lalima G.;Shaw, Andrey S.
通讯作者：
Shaw, Andrey S.
DOI：
10.1073/pnas.0914157107
发表时间：
2010-02-16
影响因子：
11.1
作者：
Bae, Jae Hyun;Boggon, Titus J.;Schlessinger, Joseph
通讯作者：
Schlessinger, Joseph
DOI：
10.1073/pnas.0812413106
发表时间：
2009-02-03
影响因子：
11.1
作者：
Gajiwala, Ketan S.;Wu, Joe C.;Demetri, George D.
通讯作者：
Demetri, George D.