PUBLICATION

cDNA cloning, expression, and functional characterization of a zebrafish SULT1 cytosolic sulfotransferase

Authors
Sugahara, T., Liu, C.-C., Carter, G., Govind, Pai, T., and Liu, M.-C.
ID
ZDB-PUB-030527-15
Date
2003
Source
Archives of biochemistry and biophysics   14(1): 67-73 (Journal)
Registered Authors
Keywords
none
MeSH Terms
  • Cloning, Molecular/methods
  • Escherichia coli/enzymology
  • Escherichia coli/genetics
  • Escherichia coli/isolation & purification
  • Temperature
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • Metals/chemistry
  • Animals
  • Zebrafish/genetics*
  • Zebrafish/metabolism*
  • Enzyme Stability
  • Recombinant Fusion Proteins/biosynthesis
  • Recombinant Fusion Proteins/chemistry
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/isolation & purification
  • Cations, Divalent/chemistry
  • Sulfotransferases/chemistry
  • Sulfotransferases/genetics*
  • Sulfotransferases/isolation & purification
  • Sulfotransferases/metabolism*
  • Enzyme Activation
  • DNA, Complementary/genetics*
  • Cytosol/enzymology
  • Cytosol/metabolism
  • Gene Expression Regulation, Enzymologic
  • Base Sequence
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
PubMed
12745256 Full text @ Arch. Biochem. Biophys.
Abstract
Using the reverse transcriptase-polymerase chain reaction technique, a full-length cDNA encoding a novel zebrafish sulfotransferase was cloned and sequenced. Sequence analysis indicated that this zebrafish sulfotransferase belongs to the SULT1 cytosolic sulfotransferase gene family. The recombinant form of the zebrafish sulfotransferase, purified from Escherichia coli cells, displayed sulfating activities toward a number of endogenous compounds, in particular dopamine and thyroid hormones, in addition to xenobiotics including some flavonoids, isoflavonoids, and other phenolic compounds. The zebrafish sulfotransferase exhibited substrate dependence in pH optimum. In comparison with those determined with dopamine as substrate, the zebrafish sulfotransferase displayed much lower K(m) and higher V(max) with n-propyl gallate as substrate. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 and 43 degrees C. Among 10 divalent metal cations tested, Hg(2+), Co(2+), Zn(2+), Cd(2+), Cu(2+), and Pb(2+) exhibited dramatic inhibitory effects on the activity of the zebrafish sulfotransferase.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping