Characterization and comparative studies of zebrafish and human recombinant dihydrofolate reductases - Inhibition by folic acid and polyphenols

Tseng Ting Kao, Kuan Chieh Wang, Wen Ni Chang, Chia Ying Lin, Bing Hung Chen, Hua Lin Wu, Guey Yueh Shi, Jen Ning Tsai, Tzu Fun Fu

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Dihydrofolate reductase (DHFR) catalyzes folic acid reduction and recycles dihydrofolate generated during dTMP biosynthesis to tetrahydrofolate. DHFR is the main target of methotrexate, the most widely used agent for antifolate therapy. Nevertheless, the emergence of methotrexate-resistance has greatly impeded the curative potential of this drug. Therefore, drugs with improved efficacy are still in demand, as well as an efficient in vitro assay system and animal model for antifolate drug discovery. The aim of this study is to evaluate the suitability of using zebrafish DHFR as an alternative assay system for antifolate drug discovery. The cDNAs encoding zebrafish and human DHFR were cloned, overexpressed, and purified. Similar structural and kinetic properties were revealed between zebrafish and human recombinant DHFRs. The susceptibilities of both enzymes to known DHFR inhibitors, including methotrexate and trimethoprim, and compounds with antifolate potential, such as polyphenols, are also comparable. In addition, the DHFR-mediated dihydrofolate reduction was significantly inhibited by its own substrate folic acid. An unexpected tissuespecific distribution of DHFR was observed with the highest level present in ova and brains of zebrafish. DHFR is also abundant in zebrafish embryos of early stages and decreased abruptly after 3 days postfertilization. The substantial resemblance between zebrafish and human DHFRs, as demonstrated in this study, provides compelling evidence supporting the use of zebrafish DHFR as an in vitro assay system for folate-related studies and drug discovery.

Original languageEnglish
Pages (from-to)508-516
Number of pages9
JournalDrug Metabolism and Disposition
Volume36
Issue number3
DOIs
Publication statusPublished - 2008 Mar

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

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