Zhu Jin-Yi, Cuellar Rebecca A, Berndt Norbert, Lee Hee Eun, Olesen Sanne H, Martin Mathew P, Jensen Jeffrey T, Georg Gunda I, Schönbrunn Ernst
Drug Discovery Department, Moffitt Cancer Center , Tampa, Florida 33612, United States.
Department of Medicinal Chemistry, University of Minnesota , Minneapolis, Minnesota 55414, United States.
J Med Chem. 2017 Sep 28;60(18):7863-7875. doi: 10.1021/acs.jmedchem.7b00996. Epub 2017 Sep 14.
Members of the Wee family of kinases negatively regulate the cell cycle via phosphorylation of CDK1 and are considered potential drug targets. Herein, we investigated the structure-function relationship of human Wee1, Wee2, and Myt1 (PKMYT1). Purified recombinant full-length proteins and kinase domain constructs differed substantially in phosphorylation states and catalytic competency, suggesting complex mechanisms of activation. A series of crystal structures reveal unique features that distinguish Wee1 and Wee2 from Myt1 and establish the structural basis of differential inhibition by the widely used Wee1 inhibitor MK-1775. Kinome profiling and cellular studies demonstrate that, in addition to Wee1 and Wee2, MK-1775 is an equally potent inhibitor of the polo-like kinase PLK1. Several previously unrecognized inhibitors of Wee kinases were discovered and characterized. Combined, the data provide a comprehensive view on the catalytic and structural properties of Wee kinases and a framework for the rational design of novel inhibitors thereof.
Wee激酶家族成员通过对细胞周期蛋白依赖性激酶1（CDK1）进行磷酸化来负调控细胞周期，被视为潜在的药物靶点。在此，我们研究了人类Wee1、Wee2和Myt1（PKMYT1）的结构-功能关系。纯化的重组全长蛋白和激酶结构域构建体在磷酸化状态和催化能力上存在显著差异，提示其激活机制复杂。一系列晶体结构揭示了Wee1和Wee2与Myt1不同的独特特征，并确立了广泛使用的Wee1抑制剂MK-1775产生差异抑制作用的结构基础。激酶组分析和细胞研究表明，除了Wee1和Wee2外，MK-1775还是polo样激酶PLK1的强效抑制剂。我们发现并表征了几种先前未被认识的Wee激酶抑制剂。综合来看，这些数据提供了关于Wee激酶催化和结构特性的全面视图，以及合理设计其新型抑制剂的框架。
