Gallastegui Edurne, Bachs Oriol
Department of Cell Biology, Immunology and Neurosciences, University of Barcelona, Barcelona, Spain.
Methods Mol Biol. 2016;1336:9-12. doi: 10.1007/978-1-4939-2926-9_2.
Cyclin-dependent kinases (Cdks) belong to a family of key regulators of cell division cycle and transcription. Their activity is mainly regulated by association with regulatory subunits named cyclins but their activities are also regulated by phosphorylation, acetylation, and the association with specific inhibitory proteins (CKIs). The activity of different Cdks is deregulated in many different type of tumors, and thus, Cdks are considered targets for antitumoral therapy. For large screenings of inhibitors the use of purified recombinant Cdks and cyclins is recommended. We report here the current methods to determine their in vitro activity for large screenings of inhibitors.
细胞周期蛋白依赖性激酶(Cdks)属于细胞分裂周期和转录的关键调节因子家族。它们的活性主要通过与名为细胞周期蛋白的调节亚基结合来调节,但其活性也受磷酸化、乙酰化以及与特定抑制蛋白(CKIs)结合的调节。在许多不同类型的肿瘤中,不同Cdk的活性失调,因此,Cdk被认为是抗肿瘤治疗的靶点。对于抑制剂的大规模筛选,建议使用纯化的重组Cdk和细胞周期蛋白。我们在此报告用于大规模筛选抑制剂的测定其体外活性的当前方法。