Identification of a novel estrogen-sulfating cytosolic SULT from zebrafish: molecular cloning, expression, characterization, and ontogeny study.
By searching the expressed sequence tag database, a zebrafish cDNA encoding a putative cytosolic sulfotransferase (SULT) was identified. Sequence analysis indicated that this zebrafish SULT belongs to the SULT1 cytosolic SULT gene family. The recombinant form of this novel zebrafish SULT, expressed using the pGEX-2TK expression system and purified from transformed BL21 (DE3) Escherichia coli cells, displayed sulfating activities specifically for estrone and 17beta-estradiol among various endogenous compounds tested as substrates. The enzyme also exhibited sulfating activities toward some xenobiotic phenolic compounds. This new zebrafish SULT showed dual pH optima, at 6.5 and 10-10.5, with estrone or n-propyl gallate as substrate. Kinetic constants of the sulfation of estrone, 17beta-estradiol, and n-propyl gallate were determined. Developmental stage-dependent expression experiments revealed a significant level of expression of this novel zebrafish estrogen-sulfating SULT at the beginning of the hatching period during embryogenesis, which continued throughout the larval stage onto maturity.
Yasuda
Shin
S
Biomedical Research Center, The University of Texas Health Center, Tyler, TX 75708, USA.
Liu
Chau-Ching
CC
Takahashi
Saki
S
Suiko
Masahito
M
Chen
Lanzhuang
L
Snow
Rhodora
R
Liu
Ming-Cheh
MC
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004967:Estrogens / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013431:Sulfates / Q000378:metabolism*; D015238:Sulfotransferases / Q000378:metabolism*; D015027:Zebrafish
D017931:DNA Primers; D004967:Estrogens; D013431:Sulfates; D015238:Sulfotransferases