Solon April, Ohi Ryoma
Department of Cell and Developmental Biology, University of Michigan School of Medicine, Ann Arbor, MI, USA.
Methods Mol Biol. 2022;2415:151-165. doi: 10.1007/978-1-0716-1904-9_11.
Mitotic kinesins play essential roles during mitotic spindle assembly and in ensuring proper chromosome segregation. Chemical inhibitors of mitotic kinesins are therefore valuable tools to study kinesin function in vitro and in cells. Because cancer is a disease of unregulated cell division, inhibitors also represent potential chemotherapeutic agents. Here, we present assays that can be used to evaluate the potency and specificity of mitotic kinesin inhibitors identified from high-throughput screening. By evaluating their effects in a variety of in vitro, fixed-cell, and live cell assays, screening hits can be prioritized and optimized to produce effective, on-target inhibitors.
有丝分裂驱动蛋白在有丝分裂纺锤体组装以及确保染色体正确分离过程中发挥着至关重要的作用。因此，有丝分裂驱动蛋白的化学抑制剂是研究驱动蛋白在体外和细胞内功能的宝贵工具。由于癌症是一种细胞分裂失控的疾病，抑制剂也代表着潜在的化疗药物。在此，我们介绍了一些检测方法，可用于评估从高通量筛选中鉴定出的有丝分裂驱动蛋白抑制剂的效力和特异性。通过在各种体外、固定细胞和活细胞检测中评估它们的作用，可以对筛选出的活性化合物进行排序和优化，以生产出有效的、作用于靶点的抑制剂。
