PUBLICATION

Identification of a novel thyroid hormone-sulfating cytosolic sulfotransferase, SULT1 ST5, from zebrafish

Authors
Yasuda, S., Kumar, A.P., Liu, M.Y., Sakakibara, Y., Suiko, M., Chen, L., and Liu, M.C.
ID
ZDB-PUB-050803-1
Date
2005
Source
The FEBS journal   272(15): 3828-3837 (Journal)
Registered Authors
Keywords
molecular cloning, sulfotransferase, SULT1, thyroid hormone, zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Arylsulfotransferase/genetics
  • Arylsulfotransferase/isolation & purification
  • Arylsulfotransferase/metabolism*
  • Cations, Divalent/metabolism
  • Cloning, Molecular
  • Dogs
  • Humans
  • Hydrogen-Ion Concentration
  • Isoenzymes/genetics
  • Isoenzymes/isolation & purification
  • Isoenzymes/metabolism*
  • Metals/metabolism
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Substrate Specificity
  • Thyroid Hormones/metabolism*
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/isolation & purification
  • Zebrafish Proteins/metabolism*
PubMed
16045754 Full text @ FEBS J.
Abstract
By employing RT-PCR in conjunction with 3'-RACE, a full-length cDNA encoding a novel zebrafish cytosolic sulfotransferase (SULT) was cloned and sequenced. Sequence analysis revealed that this zebrafish SULT (designated SULT1 ST5) is, at the amino acid sequence level, close to 50% identical to human and dog SULT1B1 (thyroid hormone SULT). A recombinant form of zebrafish SULT1 ST5 was expressed using the pGEX-2TK bacterial expression system and purified from transformed BL21 (DE3) cells. Purified zebrafish SULT1 ST5 migrated as a 34 kDa protein and displayed substrate specificity for thyroid hormones and their metabolites among various endogenous compounds tested. The enzyme also exhibited sulfating activities toward some xenobiotic phenolic compounds. Its pH optima were 6.0 and 9.0 with 3,3',5-triiodo-l-thyronine (l-T(3)) as substrate and 6.0 with beta-naphthol as substrate. Kinetic constants of the enzyme with thyroid hormones and their metabolites as substrates were determined. Quantitative evaluation of the regulatory effects of divalent metal cations on the l-T(3)-sulfating activity of SULT1 ST5 revealed that Fe(2+), Hg(2+), Co(2+), Zn(2+), Cu(2+), Cd(2+) and Pb(2+) exhibited dramatic inhibitory effects, whereas Mn(2+) showed a significant stimulation. Developmental stage-dependent expression experiments revealed a significant level of expression of this novel zebrafish thyroid hormone-sulfating SULT at the beginning of the hatching period during embryogenesis, which gradually increased to a high level of expression throughout the larval stage into maturity.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping