SRC, A PROTEIN KINASE ACTIVE IN CHRONIC MYELOID LEUKEMIA

SRC，一种在慢性粒细胞白血病中活跃的蛋白激酶

• 批准号: 8363361
• 负责人: Markus A Seeliger
• 金额: $ 0.21万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363361
• 关键词: Address; Affect; Binding; Biochemical; Biological Assay; Cell division; Chimeric Proteins; Chronic Myeloid Leukemia; Clinical; Complex; Crystallography; Cyclic AMP-Dependent Protein Kinases; Disease; Drug Delivery Systems; Drug resistance; Elements; Energy Metabolism; Event; Funding; Gleevec; Goals; Grant; Imatinib; Kinetics; Ligand Binding; Light; Mediating; Molecular Conformation; Motion; National Center for Research Resources; Nuclear Magnetic Resonance; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphotransferases; Principal Investigator; Process; Protein Kinase; Proteins; Proto-Oncogenes; Regulation; Research; Research Infrastructure; Resources; Roentgen Rays; Role; Signal Transduction; Source; Specificity; Structure; Synchrotrons; United States National Institutes of Health; Vertebral column; cost; flexibility; inhibitor/antagonist; kinase inhibitor; research study; resistance mutation; src-Family Kinases; success

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Protein kinases mediate many cell signaling events, and their tight control is essential for regulating vital processes ranging from cell division to energy metabolism. Thus, it is not surprising that protein kinases are directly or indirectly involved in many diseases and that kinases are key drug targets. For example, Src kinase was the first identified proto-oncogene and the formation of a de-regulated Abl fusion protein (BCRAbl) is the cause of disease in 95% of patients with chronic myeloid leukemia. X-ray crystal structures have shown that the same kinases can attain an active and various inactive conformations, implying that kinases are inherently flexible. How the active and inactive states are stabilized and how these states interconvert are key questions in understanding kinase regulation. Because X-ray crystal structures provide only static snapshots, we will use nuclear magnetic resonance (NMR) experiments and ligand binding kinetics to study the timescales and amplitudes of structural interconversions in Abl and Src kinase domains. BCR-Abl is the target of the clinically highly successful drug imatinib (Gleevec¿, Novartis) in the treatment of chronic myelogenous leukemia (CML). The clinical success of imatinib is due to its excellent specificity, binding only to the inactive conformation of the kinase. Therefore drug binding is intimately related to the interconversion between active and inactive states. The goal of this study is to examine timescales and pathways of these interconversions between active and inactive conformations, how dynamics of structural elements relate to catalytic turnover of the kinase and how drug resistance mutations affect these dynamics. Therefore, we will compare the timescales and amplitudes of backbone motions between Src and Abl kinases in the presence of drugs by NMR experiments. Ligand binding kinetics will be used to address the role of the regulatory domains on kinase dynamics and the binding mechanisms of different classes of kinase inhibitors. The role of protein plasticity and dynamics on inhibitor promiscuity will be addressed by structural studies on kinase+inhibitor complexes, inhibitor binding kinetics and biochemical assays.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 蛋白激酶介导许多细胞信号传导事件，并且它们的严格控制对于调节从细胞分裂到能量代谢的重要过程是必不可少的。因此，蛋白激酶直接或间接参与许多疾病并且激酶是关键药物靶标并不奇怪。例如，Src激酶是第一个被鉴定的原癌基因，并且失调的Abl融合蛋白（BCRAbl）的形成是95%的慢性髓性白血病患者的病因。 X射线晶体结构表明，相同的激酶可以获得活性和各种非活性构象，这意味着激酶是固有的灵活性。活性和非活性状态如何稳定以及这些状态如何相互转换是理解激酶调节的关键问题。由于X射线晶体结构仅提供静态快照，我们将使用核磁共振（NMR）实验和配体结合动力学来研究Abl和Src激酶结构域中结构相互转换的时间尺度和幅度。BCR-Abl是临床上非常成功的药物伊马替尼（格列卫，诺华）在治疗慢性粒细胞白血病（CML）的目标。伊马替尼的临床成功是由于其出色的特异性，仅结合激酶的非活性构象。因此，药物结合与活性和非活性状态之间的相互转换密切相关。 本研究的目标是研究活性和非活性构象之间相互转化的时间尺度和途径，结构元件的动态如何与激酶的催化转换相关，以及耐药突变如何影响这些动态。因此，我们将通过核磁共振实验比较药物存在下Src和Abl激酶骨架运动的时间尺度和幅度。配体结合动力学将用于解决激酶动力学的调节结构域的作用和不同类别的激酶抑制剂的结合机制。蛋白质的可塑性和动力学抑制剂混杂的作用将解决激酶+抑制剂复合物，抑制剂结合动力学和生化分析的结构研究。