The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Department of Medical Biology, University of Melbourne, Parkville, VIC, Australia.
Commun Biol. 2021 Sep 20;4(1):1105. doi: 10.1038/s42003-021-02631-y.
Doublecortin-like kinase 1 (DCLK1) is an understudied bi-functional kinase with a proven role in tumour growth and development. However, the presence of tissue-specific spliced DCLK1 isoforms with distinct biological functions have challenged the development of effective strategies to understand the role of DCLK1 in oncogenesis. Recently, DCLK1-IN-1 was reported as a highly selective DCLK1 inhibitor, a powerful tool to dissect DCLK1 biological functions. Here, we report the crystal structures of DCLK1 kinase domain in complex with DCLK1-IN-1 and its precursors. Combined, our data rationalises the structure-activity relationship that informed the development of DCLK1-IN-1 and provides the basis for the high selectivity of DCLK1-IN-1, with DCLK1-IN-1 inducing a drastic conformational change of the ATP binding site. We demonstrate that DCLK1-IN-1 binds DCLK1 long isoforms but does not prevent DCLK1's Microtubule-Associated Protein (MAP) function. Together, our work provides an invaluable structural platform to further the design of isoform-specific DCLK1 modulators for therapeutic intervention.
双皮质醇激酶 1（DCLK1）是一种研究不足的双功能激酶，其在肿瘤生长和发展中具有明确的作用。然而，具有不同生物学功能的组织特异性剪接 DCLK1 同工型的存在，给理解 DCLK1 在肿瘤发生中的作用的有效策略的发展带来了挑战。最近，DCLK1-IN-1 被报道为一种高度选择性的 DCLK1 抑制剂，是解析 DCLK1 生物学功能的有力工具。在这里，我们报告了 DCLK1 激酶结构域与 DCLK1-IN-1 及其前体复合物的晶体结构。综合我们的数据，合理化了结构-活性关系，为 DCLK1-IN-1 的发展提供了基础，并解释了 DCLK1-IN-1 的高选择性，DCLK1-IN-1 诱导 ATP 结合位点的剧烈构象变化。我们证明 DCLK1-IN-1 结合 DCLK1 长同工型，但不阻止 DCLK1 的微管相关蛋白（MAP）功能。总之，我们的工作提供了一个宝贵的结构平台，以进一步设计针对治疗干预的同工型特异性 DCLK1 调节剂。
