体外反应体系中APC/C活性及相关研究

Activity of APC/C and Its Relationship with the MCC in the In vitro Reaction System

为利用体外反应体系研究纯化的细胞周期后期促进复合物(APC/C)相关的分子事件,以其底物泛素化为检测指标,分别从来源细胞裂解液浓度,不同底物间泛素化强度,以及与纺锤体检查点复合物的解离等几方面进行了研究。结果显示,由浓度高的细胞裂解液纯化出的APC/C活性高,其对底物Geminin的泛素化活性较Cyclin B高,在APC/C活性由被抑制到释放过程中,APC/C与Mad2,BubR1结合减少且Cdc20泛素化增强。

To analysis the activity of APC/C and its relationship with the MCC in the In vitro reaction system,through detecting the substrates＇ ubiquitination,several assays were done.The concentration of extracts where purified APC/C from,different substrates,and the disassociation of MCC from APC/C were analyzed.As shown in the results,the activity of APC/C purified from the higher concentration extracts was higher,the ubiquitination of Geminin was easier than the Cyclin B and during the activation of APC/C,the binding of Mad2 and BubR1 was decreased,while the ubiquitination of Cdc20 was increased.