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Targeting differential kinase domain dimerization of EGFR mutants

Targeting differential kinase domain dimerization of EGFR mutants
靶向 EGFR 突变体的差异激酶结构域二聚化
批准号:
10201967
负责人:
Yuko Tsutsui
金额:
$16.75万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2021
资助国家:
美国
项目状态:
已结题
起止时间:
2021-05-01 至 2023-04-30

项目摘要

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中文摘要
翻译
在这个R03应用程序中,PI将在Lemmon实验室开发一个旨在理解的独立项目
英文摘要
In this R03 application, the PI will develop an independent project in the Lemmon lab aimed at understanding the relative importance of mutation-induced dimerization (versus direct activation) of the kinase domain of the Epidermal Growth Factor Receptor (EGFR) in lung cancer patients. The EGFR regulates critical cellular processes such as cell motility, metabolism, proliferation and differentiation, and is normally activated by ligand- induced dimerization. In a subset of lung cancer patients, kinase domain mutations activate EGFR in the absence of ligand. Traditionally, these mutations (such as L858R and exon 19 deletions) have been thought to bias the monomeric kinase domain to its active state – independent of dimerization. However, some recent studies suggest that kinase domain dimerization is key to mutational activation of EGFR. Moreover, several rare EGFR mutations – such as those in exon 18, or which cause kinase domain duplication (KDD) – seem to activate EGFR primarily by dimerizing its kinase domain. In this proposal, we ask whether/how common EGFR mutations promote kinase domain dimerization, and whether they are capable of ‘super-activating’ a kinase domain dimer. In addition, we address the ‘dimerization only’ activation mechanisms of exon 18 and KDD mutations. By understanding these different modes of activation, we hope to gain insight into new therapeutic approaches to inhibit EGFR by targeting the dimerization interface – which could be mutant-specific in detail. Through in vitro kinase assay approaches to establish reaction kinetics, as well as a host of biophysical methods, structural analysis, and computational tools (validated experimentally) we propose three Specific Aims: Aim 1: Understanding relative contribution of kinase domain dimerization to activation of EGFR by oncogenic mutations. Using a synthetic disulfide-linked EGFR kinase domain dimer, we will study the elusive asymmetric EGFR kinase domain dimer biochemically. We will ask whether common EGFR mutations further enhance activity of this dimer, and will compare activities of dimers induced by exon 18 and KDD mutations. Aim 2: Structural analysis and ‘in-solution’ dynamics of the EGFR kinase domain harboring exon 18 mutations or kinase domain duplications (KDD) that promote ligand-independent dimerization. Protein crystallography will be pursued to visualize the interfaces of the different kinase domain dimers – to ask whether they are identical or mutant-specific. In parallel, hydrogen-deuterium exchange mass spectrometry (HDX-MS) will be employed to map interfaces and their stability. These studies will be complemented by other biophysical studies to gain a view of dimer stability and dynamics for dimers induced by different means and mutations. Aim 3: Targeting the kinase domain dimerization interface by small molecules as disruptors of protein- protein interactions (PPIs). Finally, we will identify structural motifs that can bind to the EGFR kinase domain dimerization interface and disrupt allosteric activation in an uncompetitive or non-competitive fashion by screening libraries of macrocyclic small molecules in the Yale Center for Molecular Discovery.
期刊论文(1)
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科研奖励(0)
会议论文
DOI: 10.1016/bs.mie.2022.03.037
发表时间: 2022
期刊: Methods in enzymology
影响因子: --
作者: []
通讯作者:
国内基金
海外基金
帽结合蛋白(cap binding protein)调控乙烯信号转导的分子机制
  • 批准号:
    32170319
  • 项目类别:
    面上项目
  • 资助金额:
    58.00万元
  • 批准年份:
    2021
  • 负责人:
    董春海
  • 依托单位:
帽结合蛋白(cap binding protein)调控乙烯信号转导的分子机制
  • 批准号:
    --
  • 项目类别:
    --
  • 资助金额:
    58万元
  • 批准年份:
    2021
  • 负责人:
    董春海
  • 依托单位:
ID1 (Inhibitor of DNA binding 1) 在口蹄疫病毒感染中作用机制的研究
番茄EIN3-binding F-box蛋白2超表达诱导单性结实和果实成熟异常的机制研究
  • 批准号:
    31372080
  • 项目类别:
    面上项目
  • 资助金额:
    80.0万元
  • 批准年份:
    2013
  • 负责人:
    杨迎伍
  • 依托单位: