Activation and Regulation Mechanisms of the RAF Kinase Family

BRAF是RAF家族蛋白激酶的一员，参与MAPK信号通路，控制BREAD 细胞活动，如增殖和分化。BRAF是人类最常见的突变蛋白 癌症。此外，当有突变时，缺乏BRAF突变的肿瘤取决于BRAF的活性 在上游通路成员中，如RAS或生长因子受体。因此，BRAF代表着一个重要的 癌症治疗的靶点。尽管经过了三十年的紧张研究，直到最近才有了足够的了解 对BRAF机制的研究为BRAF治疗打开了大门。然而，目前临床上使用的BRAF抑制剂， 维莫拉非尼和达普拉非尼仅限于I类突变体BRAFV600E。维莫拉非尼布和达普拉非尼布，两人 ATP竞争性抑制剂可以矛盾地激活非BRAFV600E和野生型BRAF，这表明 BRAF具有独立于其激酶活性的功能。非小细胞肺癌的治疗策略 BRAFV600E突变和野生型BRAF仍然缺乏。我们知识上的差距导致了这些局限性 三磷酸腺苷-竞争性BRAF药物。拟议的工作围绕着关于英国皇家空军激活的三个问题 以及健康和疾病状态的监管。问题1：BRAF结构元素和ATP结合如何调节 非BRAFV600E和野生型BRAF？问2：红花的作用机制是什么？ 本课题组开发的第一个BRAF变构抑制剂以及如何靶向催化和非催化的 BRAF的催化功能有待开发？问题3：英国皇家空军职能不重叠的原因是什么？ 异构体，尽管它们的结构相似？私家侦探和她的团队将通过 多学科方法，包括X射线结晶学、结合动力学、磷酸蛋白质组学、活细胞 成像、化学生物学和计算方法。我们的发现不仅有助于更好地理解 RAF激酶家族复杂的生化机制，也为新的 针对BRAF驱动的肿瘤的治疗方法。

BRAF, a member of the RAF family protein kinases, works in the MAPK signaling pathway, which controls broad cellular events like proliferation and differentiation. BRAF is the most frequently mutated kinase in human cancers. Furthermore, tumors lacking BRAF mutations are contingent on BRAF activity when there are mutations in upstream pathway members, such as RAS or growth factor receptors. Thus, BRAF represents an important target for cancer therapy. Despite three decades of intense research, only recently has a sufficient understanding of BRAF's mechanism creaked open the door to BRAF therapy. However, the current clinical BRAF inhibitors, vemurafenib and dabrafenib, are limited to the class I mutant BRAFV600E. Vemurafenib and dabrafenib, the two ATP-competitive inhibitors, can paradoxically activate non-BRAFV600E and wild-type BRAF, suggesting that BRAF has functions that are independent of its kinase activity. Therapeutic strategies for tumors with non- BRAFV600E mutations and wild-type BRAF are still lacking. Gaps in our knowledge contributed to the limitations of ATP-competitive BRAF drugs. The proposed work centers around three questions regarding RAF activation and regulation in health and disease states. Q1: how BRAF structural elements and ATP binding mediate the catalytic-independent functions of non-BRAFV600E and wild-type BRAF? Q2: what is the mechanism of action of the first BRAF allosteric inhibitor developed by our group and how can inhibitors targeting the catalytic and non- catalytic functions of BRAF be developed? Q3: What factors contribute to the non-overlapping functions of RAF isoforms, despite their structural similarity? The PI and her team will address these questions through multidisciplinary approaches, including X-ray crystallography, binding kinetics, phosphoproteomics, live cell imaging, chemical biology, and computational methods. Our findings will not only facilitate a better understanding of the complex biochemical mechanisms of the RAF kinase family, also provide a molecular basis for novel therapeutic approaches targeting BRAF-driven tumors.