As a member of the Wee-kinase family protein kinase PKMYT1 is involved in G₂/M checkpoint regulation of the cell cycle. Recently, a peptide microarray approach led to the identification of a small peptide; EFS^247–259 as substrate of PKMYT1, which allowed for subsequent development of an activity assay. The developed activity assay was used to characterize the PKMYT1 catalyzed phosphorylation of EFS^247–259. For the first time kinetic parameters for PKMYT1, namely K_m, K_{m, ATP} and v_max were determined. The optimized assay was used to screen the published protein kinase inhibitor sets (PKIS I and II), two sets of small molecule ATP-competitive kinase inhibitors reported by GlaxoSmithKline. We identified ten inhibitors, providing different scaffolds. The inhibitors were further characterized by using binding assay, activity and functional assay. In addition, docking studies were carried out in order to rationalize the observed biological activities. The derived results provide the basis for further chemical optimization of PKMYT1 inhibitors and for further analysis of PKMYT1 as target for anti-cancer therapy.

作为Wee激酶家族的成员，蛋白激酶PKMYT1参与细胞周期的G ₂ / M检查点调节。最近，一种肽微阵列方法导致了对小肽的鉴定。EFS ^247-259作为^PKMYT1的底物，可用于后续的活性测定。发达的活性测定法用于表征PKMYT1催化EFS ^247–259的磷酸化。PKMYT1的动力学参数首次为K _m，K _m，ATP和v _max被确定。优化的分析方法用于筛选已发布的蛋白激酶抑制剂组（PKIS I和II），这是GlaxoSmithKline报告的两组小分子ATP竞争性激酶抑制剂。我们确定了十种抑制剂，提供了不同的支架。通过使用结合测定，活性和功能测定来进一步表征抑制剂。另外，进行了对接研究以使观察到的生物活性合理化。得出的结果为进一步优化PKMYT1抑制剂的化学性质和进一步分析PKMYT1作为抗癌治疗的靶标奠定了基础。