Structure

项目总结 最近发现的导致非小细胞肺癌的基因改变为治疗干预提供了潜在的有吸引力的靶点。在这个项目中，我们关注三个这样的靶点，它们都是激酶：表皮生长因子受体(EGFR)、坦克结合激酶1(TBK1)和盘状结构域受体2(DDR2)的耐药突变。明确的结构和机制了解致癌突变对激酶活性和抑制剂敏感性的影响，对于高效和有效地开发新的抑制剂是至关重要的。此外，当受到先导化合物的共晶结构的启发时，缓蚀剂的发现和优化要高效得多。蛋白质结构和酶学核心将通过提供纯化的重组激酶、酶和抑制剂表征、分子建模以及突变激酶和激酶/抑制剂复合体的晶体结构测定方面的专业知识，为这三个项目的每一个项目提供支持。将这些活动集中在核心将防止在各个项目中重复工作和专业知识，并将利用环境保护与可持续发展实验室现有的资源和经验。我们在结构生物学和激酶抑制方面拥有丰富的经验，并与项目负责人和核心主管建立了长期的合作关系。这些项目(以及与核心主任实验室中正在进行的其他项目)之间的协同作用以及“分阶段”启动的结晶工作将使我们能够以相对较少的资源实现我们的目标。 在每个项目和化学核心的密切合作下，我们将完成以下具体目标：1)EGFR激酶耐药突变的结晶学和机制研究；2)TBK1的表达、纯化和结构测定；以及3)DDR2酪氨酸激酶癌基因突变的结构和机制表征。

PROJECT SUMMARY Recently identified genetic alterations that drive non-small cell lung cancer provide potentially attractive targets for therapeutic intervention. In this Program, we focus on three such targets, all of which are kinases: drug-resistant mutants of the epidermal growth factor receptor (EGFR), TANK binding kinase 1 (TBKl) and discoidin domain receptor 2 (DDR2). A clear structural and mechanistic understanding of the effects of oncogenic mutations on the activity of a kinase and on inhibitor sensitivity is essential for efficient and effective development of new inhibitors. Additionally, inhibitor discovery and optimization is much more efficient when illuminated by co-crystal structures of lead compounds. The Protein Structure and Enzymology core will support each of the three projects by providing purified recombinant kinases, expertise in enzyme and inhibitor characterization, molecular modeling, and crystallographic structure determination of mutant kinases and kinase/inhibitor complexes. Centralizing these activities in the core will prevent duplication of effort and expertise in the individual projects and will leverage the resources and experience available in the Eck laboratory. We have extensive experience in the structural biology and inhibition of kinases and long-standing collaborative ties with the Project Leaders and Core Directors. Synergies between the projects (and with other projects ongoing in the Core Director's laboratory) and "staged" initiation of crystallographic efforts will allow us to accomplish our aims with relatively modest resources. In close collaboration with each of the Projects and with the Chemistry Core, we will accomplish the following specific aims: 1) Crystallographic and mechanistic studies of drug-resistant mutants of the EGFR kinase 2) Expression, purification and structure determination of TBK1, and 3) Structural and mechanistic characterization of oncogenic mutants of the DDR2 tyrosine kinase.