Conformational Dynamics of Protein Tyrosine Kinases Src and Abl

蛋白激酶介导许多细胞信号事件，它们的严密控制对调节生命至关重要。 从细胞分裂到能量新陈代谢的过程。因此，并不奇怪，蛋白激酶是 直接或间接地与许多疾病有关，而激酶是关键的药物靶标。例如，源 激酶是第一个被发现的原癌基因和去调控的abl融合蛋白的形成。 (BCRAbI)是95%的慢性髓系白血病患者的致病原因。X射线晶体结构 已经表明，相同的激酶可以获得活跃的和各种不活跃的构象，这意味着 肌动蛋白具有天生的灵活性。活动状态和非活动状态如何稳定以及这些状态如何稳定 相互转换是理解激酶调节的关键问题。因为X射线晶体结构提供了 只有静态快照，我们将使用核磁共振(核磁共振)实验和配体结合 研究Abl和Src结构域结构转换的时间尺度和幅度的动力学。 Bcr-abi是临床上非常成功的药物伊马替尼(格列卫，诺华)治疗糖尿病的靶点。 慢性粒细胞白血病(CML)。伊马替尼的临床成功归功于其出色的特异性， 只与非活性构象的激酶结合。因此，药物结合与生物多样性密切相关。 活动状态和非活动状态之间的相互转换。这项研究的目标是检查时间尺度和 这些活动构象和非活动构象之间相互转化的途径，结构的动力学 这些元素与激酶的催化周转以及耐药突变如何影响这些动态有关。 因此，我们将比较src和abl的脊椎运动的时间尺度和幅度。 在药物存在的情况下进行核磁共振实验。配体结合动力学将被用来解决 调控结构域在蛋白激酶动力学中的作用及不同类型蛋白的结合机制 这是一种激酶抑制剂。蛋白质的可塑性和动力学在抑制剂混杂中的作用将通过 激酶*抑制剂复合体的结构研究、抑制剂结合动力学和生化分析。

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 (BCRAbI) 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-AbI 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 ofthese 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.