Structural studies of molecular cancer targets and drug development

目前正在研究两个系统作为校内合作。高亲和力 生长因子受体结合蛋白2（Grb 2）SH 2结构域与 受体酪氨酸激酶（RTK）的酪氨酸磷酸化胞质结构域是一种有吸引力的 在许多类型癌症中作为治疗干预的靶点。为此，我们决定 Grb 2 SH 2结构域和有效的 不含磷的大环肽模拟物， 对erbB-2依赖性乳腺癌的抗增殖作用。此代理代表一个 第二代抑制剂，结合亲和力大大提高， 生物利用度与其开链对应物相比。在2.0 ℃下测定了结构 和1.8，每个不对称单元具有一个和两个结构域交换的二聚体， 分别蛋白质和蛋白质之间的结合模式和特异性相互作用 抑制剂提供了对这类大环化合物的高效力的深入了解， 有助于作为迭代合理药物设计过程的一部分的进一步优化。我们也 参与多学科合作，开发有效和特异性的抑制剂， 人Chk 2激酶。我们在这个项目中的作用是确定Chk 2的共晶结构， 与小分子抑制剂复合。到目前为止，我们已经确定了8个共晶结构 与不同的化合物，这提供了丰富的信息模式， 抑制剂结合，并导致了关于如何提高特异性和效力的想法。我们还 产生的位点特异性生物素化的Chk 2用于表面等离子体共振研究， 抑制剂结合。最后，我们确定了人LRF的POZ结构域的结构， 肿瘤发生的主要调节因子，作为开发这种抑制剂的第一步， 蛋白质（见科学进展）。

Two systems are currently under study as intramural collaborations. The high-affinity binding of the growth-factor receptor-bound protein 2 (Grb2) SH2 domain to tyrosine-phosphorylated cytosolic domains of receptor tyrosine kinases (RTKs) is an attractive target for therapeutic intervention in many types of cancer. To this end, we have determined the structures of two crystal forms of a complex between the Grb2 SH2 domain and a potent non-phosphorus containing macrocyclic peptide mimetic that exhibits significant anti-proliferative effects against erbB-2-dependent breast cancers. This agent represents a second generation inhibitor with greatly improved binding affinity and bioavailability compared to its open-chain counterpart. The structures were determined at 2.0 and 1.8 with one and two domain-swapped dimers per asymmetric unit, respectively. The mode of binding and specific interactions between the protein and the inhibitor provide insight into the high potency of this class of macrocylic compounds and may aid in further optimization as part of the iterative rational drug design process. We are also involved in a multidisciplinary collaboration to develop potent and specific inhibitors of human Chk2 kinase. Our role in this project is to determine co-crystal structures of Chk2 in complex with small molecule inhibitors. Thus far we have determined 8 co-crystal structures with different compounds and this has provided a wealth of information about the mode of inhibitor binding and led to ideas about how to improve specificity and potency. We have also produced site-specifically biotinylated Chk2 for surface plasmon resonance studies of inhibitor binding. Finally, we have determined the structure of the POZ domain of human LRF, a master regulator of oncogenesis, as a first step toward the development of inhibitors of this protein (see Scientific Advances).