PUBLICATION
Characterization and ontogenic study of novel steroid-sulfating SULT3 sulfotransferases from zebrafish
- Authors
- Yasuda, T., Yasuda, S., Williams, F.E., Liu, M.Y., Sakakibara, Y., Bhuiyan, S., Snow, R., Carter, G., and Liu, M.C.
- ID
- ZDB-PUB-080728-3
- Date
- 2008
- Source
- Molecular and Cellular Endocrinology 294(1-2): 29-36 (Journal)
- Registered Authors
- Williams, Fred
- Keywords
- Sulfotransferase, SULT, 17β-estradiol, dehydroepiandrosterone, molecular cloning, developmental expression, zebrafish
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Cloning, Molecular
- Cytosol/enzymology
- Gene Expression Regulation, Developmental
- Hydrogen-Ion Concentration
- Kinetics
- Molecular Sequence Data
- Phylogeny
- Recombinant Proteins/genetics
- Recombinant Proteins/isolation & purification
- Recombinant Proteins/metabolism
- Steroids/metabolism*
- Substrate Specificity
- Sulfates/metabolism*
- Sulfotransferases/chemistry
- Sulfotransferases/genetics*
- Sulfotransferases/isolation & purification
- Sulfotransferases/metabolism
- Xenobiotics/metabolism
- Zebrafish/genetics*
- PubMed
- 18644423 Full text @ Mol. Cell. Endocrinol.
Citation
Yasuda, T., Yasuda, S., Williams, F.E., Liu, M.Y., Sakakibara, Y., Bhuiyan, S., Snow, R., Carter, G., and Liu, M.C. (2008) Characterization and ontogenic study of novel steroid-sulfating SULT3 sulfotransferases from zebrafish. Molecular and Cellular Endocrinology. 294(1-2):29-36.
Abstract
In vertebrates, sulfation as catalyzed by members of the cytosolic sulfotransferase (SULT) family has been suggested to be involved in the homeostasis of steroids. To establish the zebrafish as a model for investigating how sulfation functions to regulate steroid metabolism during the developmental process, we have embarked on the identification of steroid-sulfating SULTs in zebrafish. By searching the GenBank database, we identified two putative cytosolic SULT sequences from zebrafish, designated SULT3 ST1 and ST2. The recombinant proteins of these two zebrafish SULT3 STs were expressed in and purified from BL21 (DE3) cells transformed with the pGEX-2TK expression vector harboring SULT3 ST1 or ST2 cDNA. Upon enzymatic characterization, purified SULT3 ST1 displayed the strongest sulfating activity toward 17beta-estradiol among the endogenous substrates tested, while SULT3 ST2 exhibited substrate specificity toward hydroxysteroids, particularly dehydroepiandrosterone (DHEA). The pH-dependence and kinetic constants of these two enzymes with 17beta-estradiol and DHEA were determined. A developmental expression study revealed distinct patterns of the expression of SULT3 ST1 and ST2 during embryonic development and throughout the larval stage onto maturity. Collectively, these results imply that these two steroid-sulfating SULT3 STs may play differential roles in the metabolism and regulation of steroids during zebrafish development and in adulthood.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping