Expression, purification and activity assay of recombinant human thymidylate synthase  

胸苷酸合酶的表达、纯化和抑制剂筛选体系的建立(英文)

作　　者：李超[1] 郭莹[1] 田超[1] 周受辛[1] 闫汝峰[1] 张志丽[1] 王孝伟[1] 刘俊义[1] 

机构地区：[1]北京大学医学部药学院化学生物学系,北京100191

出　　处：《Journal of Chinese Pharmaceutical Sciences》2013年第2期173-176,共4页中国药学（英文版）

基　　金：supported by National Natural Science Foundation of China(Grant No.20972011,21042009,21172014)

摘　　要：Thymidylate synthase （TS, E.C.2.1.1.45） catalyzes a critical reaction in the only pathway of de novo synthesis of thymidylate （dTMP） in human cells, and is an important target of chemotherapy. To evaluate the inhibitory activities of novel compounds to TS, a convenient method of activity assay using 6x His-tagged recombinant human TS （rhTS） was established and 49 novel synthetic folate analogues were screened to discover potential TS inhibitors. During the process, 4 novel compounds were found to effectively inhibit TS, while the IC 50 of a positive control raltitrexed was 3.4 μM in this assay.胸苷酸合酶(TS)催化人体内脱氧胸苷酸唯一的从头合成反应,是抗叶酸代谢类药物的重要靶点。为方便地筛选化合物对TS的抑制活性,本论文建立了重组人TS为基础的表达、纯化和活性筛选体系,并以其为基础进行了新化合物的TS抑制活性筛选,其中4个化合物显示出了较好的抑制活性,阳性对照药物雷替曲塞的IC50值为3.4μM。实验结果表明该方法具有方便、快速和稳定的特点。

关 键 词：Thymidylate synthase inhibitors Recombinant protein expression Ni-resin column Screening model 

分 类 号：R914[医药卫生—药物化学]