PUBLICATION
Molecular cloning, expression, and functional characterization of a novel zebrafish cytosolic sulfotransferase
- Authors
- Sugahara, T., Liu, C.-C., Govind Pai, T., and Liu, M.-C.
- ID
- ZDB-PUB-030115-19
- Date
- 2003
- Source
- Biochemical and Biophysical Research Communications 300(3): 725-730 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Molecular Weight
- Substrate Specificity
- Sulfotransferases/biosynthesis
- Sulfotransferases/chemistry*
- Sulfotransferases/genetics*
- Cloning, Molecular
- Hydrogen-Ion Concentration
- Cytosol/enzymology*
- Electrophoresis, Polyacrylamide Gel
- Enzyme Activation/drug effects
- Animals
- Amino Acid Sequence
- Cations, Divalent/pharmacology
- Enzyme Stability/physiology
- Base Sequence
- Sequence Analysis, DNA
- Molecular Sequence Data
- Temperature
- Zebrafish
- Sequence Homology, Amino Acid
- Zebrafish Proteins/biosynthesis
- Zebrafish Proteins/chemistry*
- Zebrafish Proteins/genetics*
- PubMed
- 12507510 Full text @ Biochem. Biophys. Res. Commun.
Citation
Sugahara, T., Liu, C.-C., Govind Pai, T., and Liu, M.-C. (2003) Molecular cloning, expression, and functional characterization of a novel zebrafish cytosolic sulfotransferase. Biochemical and Biophysical Research Communications. 300(3):725-730.
Abstract
By searching the zebrafish expressed sequence tag (EST) database, we have identified a cDNA clone encoding a putative zebrafish cytosolic sulfotransferase (ST). This cDNA was isolated and subjected to nucleotide sequencing. Analysis of the sequence data revealed that this novel zebrafish ST displays 32-35% amino acid sequence identity to members of all major cytosolic ST gene families. Therefore, this zebrafish ST, while belonging to the cytosolic ST gene superfamily, appears to be independent from all known constituent ST gene families. Recombinant zebrafish ST, expressed using the pET23c prokaryotic expression vector and purified from transformed Escherichia coli cells, migrated as a 34-kDa protein upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified zebrafish ST displayed sulfating activities toward dopamine and thyroid hormones (T(3) and T(4)), with a pH optimum spanning 7-9. The enzyme also exhibited activities toward a number of xenobiotics including some flavonoids, isoflavonoids, and other phenolic compounds. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 and 48 degrees C. Among 10 divalent metal cations tested, Fe(++), Hg(++), Co(++), Zn(++), Cu(++), and Cd(++) exhibited dramatic inhibitory effects on the activity of the enzyme. These results constitute a first study on the cloning, expression, and characterization of a zebrafish cytosolic ST.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping