Expression, purification and activity assay of recombinant human thymidylate synthase

doi:10.5246/jcps.2013.02.024

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Expression, purification and activity assay of recombinant human thymidylate synthase

Chao Li, Ying Guo, Chao Tian, Shouxin Zhou, Rufeng Yan, Zhili Zhang, Xiaowei Wang, Junyi Liu^*

Department of Chemical Biology, School of Pharmaceutical Science, Peking University Health Science Center, Beijing 100191, China

Received:2012-12-11 Revised:2013-02-03 Online:2013-03-18 Published:2013-03-18
Contact: Junyi Liu*

Abstract

Abstract:

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 IC50 of a positive control raltitrexed was 3.4 μM in this assay.

Key words: Thymidylate synthase inhibitors, Recombinant protein expression, Ni-resin column, Screening model

CLC Number:

R916.41

Supporting:

Chao Li, Ying Guo, Chao Tian, Shouxin Zhou, Rufeng Yan, Zhili Zhang, Xiaowei Wang, Junyi Liu^*. Expression, purification and activity assay of recombinant human thymidylate synthase[J]. , DOI: 10.5246/jcps.2013.02.024.

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Expression, purification and activity assay of recombinant human thymidylate synthase

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