PUBLICATION
Characterization and comparative studies of zebrafish and human recombinant dihydrofolate reductases --- Inhibition by folic acid and polyphenols
- Authors
- Kao, T.T., Wang, K.C., Chang, W.N., Lin, C.Y., Chen, B.H., Wu, H.L., Shi, G.Y., Tsai, J.N., and Fu, T.F.
- ID
- ZDB-PUB-071210-27
- Date
- 2008
- Source
- Drug metabolism and disposition: the biological fate of chemicals 36(3): 508-516 (Journal)
- Registered Authors
- Chang, Wen-Ni, Fu, Tzu-Fun, Kao, Tseng-Ting
- Keywords
- anticancer agents, drug discovery, drug targeting, enzyme inhibitors, enzyme kinetics
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Cloning, Molecular
- DNA, Complementary/genetics
- Flavonoids/chemistry
- Flavonoids/pharmacology*
- Folic Acid/chemistry
- Folic Acid/pharmacology*
- Folic Acid Antagonists/chemistry
- Folic Acid Antagonists/pharmacology*
- Humans
- Molecular Sequence Data
- Phenols/chemistry
- Phenols/pharmacology*
- Polyphenols
- Protein Structure, Quaternary
- Protein Structure, Secondary
- Recombinant Proteins/biosynthesis
- Recombinant Proteins/chemistry
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Reverse Transcriptase Polymerase Chain Reaction
- Sequence Alignment
- Tetrahydrofolate Dehydrogenase/biosynthesis
- Tetrahydrofolate Dehydrogenase/chemistry
- Tetrahydrofolate Dehydrogenase/genetics
- Tetrahydrofolate Dehydrogenase/metabolism*
- Zebrafish
- PubMed
- 18056255 Full text @ Drug Metab. Dispos.
Citation
Kao, T.T., Wang, K.C., Chang, W.N., Lin, C.Y., Chen, B.H., Wu, H.L., Shi, G.Y., Tsai, J.N., and Fu, T.F. (2008) Characterization and comparative studies of zebrafish and human recombinant dihydrofolate reductases --- Inhibition by folic acid and polyphenols. Drug metabolism and disposition: the biological fate of chemicals. 36(3):508-516.
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 tissue-specific distribution of DHFR was observed with the highest level present in ova and brain of zebrafish. DHFR is also abundant in zebrafish embryos of early stages and decreased abruptly after 3 day-post-fertilization. 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping