PUBLICATION
Identification, characterization, and ontogenic study of a catechol O-methyltransferase from zebrafish
- Authors
- Alazizi, A., Liu, M.Y., Williams, F.E., Kurogi, K., Sakakibara, Y., Suiko, M., and Liu, M.C.
- ID
- ZDB-PUB-110317-22
- Date
- 2011
- Source
- Aquatic toxicology (Amsterdam, Netherlands) 102(1-2): 18-23 (Journal)
- Registered Authors
- Williams, Fred
- Keywords
- Catechol O-methyltransferase, Developmental expression, Methylation, Molecular cloning, Zebrafish
- MeSH Terms
-
- Animals
- Base Sequence
- Catechol O-Methyltransferase/genetics*
- Catechol O-Methyltransferase/metabolism*
- Catechols/metabolism
- Cloning, Molecular*
- Enzyme Assays
- Gene Expression
- Gene Expression Regulation, Developmental
- Metabolic Detoxication, Phase II/genetics
- Metabolic Detoxication, Phase II/physiology*
- Methylation
- Models, Animal
- Molecular Sequence Data
- Polymerase Chain Reaction
- Xenobiotics/metabolism
- Zebrafish/genetics
- Zebrafish/metabolism*
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism*
- PubMed
- 21371608 Full text @ Aquat. Toxicol.
Citation
Alazizi, A., Liu, M.Y., Williams, F.E., Kurogi, K., Sakakibara, Y., Suiko, M., and Liu, M.C. (2011) Identification, characterization, and ontogenic study of a catechol O-methyltransferase from zebrafish. Aquatic toxicology (Amsterdam, Netherlands). 102(1-2):18-23.
Abstract
To establish the zebrafish as a model for investigating the methylation pathway of drug metabolism, we embarked on the molecular cloning of the zebrafish catechol O-methyltransferase (COMT). By searching the GenBank database, a zebrafish nucleotide sequence encoding a putative COMT was identified. Based on the sequence information, we designed and synthesized oligonucleotides corresponding to its 5'- and 3'-coding regions of this zebrafish COMT. Using the first-strand cDNA reverse-transcribed from the total RNA isolated from a 3-month-old adult female zebrafish as the template, the cDNA encoding the zebrafish COMT was PCR-amplified. The recombinant zebrafish COMT protein was subsequently expressed in and purified from BL21 (DE3) Escherichia coli cells transformed with the pGEX-2TK expression vector harboring the zebrafish COMT cDNA. Upon enzymatic characterization, purified COMT displayed methylating activity toward dopamine, dopa, and catecholestrogens, as well as three representative catechol drugs, methyldopa, dobutamine, and isoproterenol. A reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed developmental stage-dependent expression of the zebrafish COMT during embryonic development and throughout the larval stage onto maturity. These results provide a foundation for investigating the involvement of COMT-mediated methylation in protection against the adverse effects of catechol drugs and other xenobiotic catechols during the developmental process.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping