title: Identification of a novel estrogen-sulfating cytosolic SULT from zebrafish: molecular cloning, expression, characterization, and ontogeny study.
abstract: 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.
authors.affiliation: Biomedical Research Center, The University of Texas Health Center, Tyler, TX 75708, USA.
authors.forename: Shin
authors.initials: S
authors.lastname: Yasuda
authors.forename: Chau-Ching
authors.initials: CC
authors.lastname: Liu
authors.forename: Saki
authors.initials: S
authors.lastname: Takahashi
authors.forename: Masahito
authors.initials: M
authors.lastname: Suiko
authors.forename: Lanzhuang
authors.initials: L
authors.lastname: Chen
authors.forename: Rhodora
authors.initials: R
authors.lastname: Snow
authors.forename: Ming-Cheh
authors.initials: MC
authors.lastname: Liu
mesh_terms: 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
chemical_list: D017931:DNA Primers; D004967:Estrogens; D013431:Sulfates; D015238:Sulfotransferases