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Sugahara T, Yang YS, Liu CC, et al. Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase. Biochem J. 2003;375:785-91doi: 10.1042/BJ20031050.
Sugahara, T., Yang, Y. S., Liu, C. C., Pai, T. G., & Liu, M. C. (2003). Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase. The Biochemical journal, 375785-91. https://doi.org/10.1042/BJ20031050
Sugahara, Takuya, et al. "Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase." The Biochemical journal vol. 375 (2003): 785-91. doi: https://doi.org/10.1042/BJ20031050
Sugahara T, Yang YS, Liu CC, Pai TG, Liu MC. Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase. Biochem J. 2003 Nov 01;375:785-91. doi: 10.1042/BJ20031050. PMID: 12885295; PMCID: PMC1223711.
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Biochem J. 2003 Nov 01;375:785-91. doi: 10.1042/BJ20031050.

Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase.

The Biochemical journal

Takuya Sugahara, Yuh-Shyong Yang, Chau-Ching Liu, T Govind Pai, Ming-Cheh Liu

Affiliations

  1. Biomedical Research Center, The University of Texas Health Center, Tyler, TX 75708, USA.

PMID: 12885295 PMCID: PMC1223711 DOI: 10.1042/BJ20031050

Abstract

By searching the zebrafish EST (expressed-sequence tag) database, we have identified two partial cDNA clones encoding the 5' and 3' regions of a putative zebrafish sulphotransferase (ST). Using the reverse transcription-PCR technique, a full-length cDNA encoding this zebrafish ST was successfully cloned. Sequence analysis revealed that this novel zebrafish ST displays 44%, 43% and 40% amino acid identity with mouse SULT2B1, human SULT2B1b and human SULT2A1 ST respectively. This zebrafish ST therefore appears to belong to the SULT2 cytosolic ST gene family. Recombinant zebrafish ST, expressed using the pGEX-2TK prokaryotic expression system and purified from transformed Escherichia coli cells, migrated as a 34 kDa protein upon SDS/PAGE. Purified zebrafish ST displayed a strong sulphonating activity toward DHEA (dehydroepiandrosterone), with a optimum pH of 9.5. The enzyme also exhibited activities toward several neurosteroids with differential K(m) and V(max) values. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 degrees C and 43 degrees C. Among ten different divalent metal cations tested, Fe2+ and Cd(2+ exhibited small, but significant, stimulatory effects, whereas Hg2+ and Cu2+ displayed considerably stronger inhibitory effects on the DHEA-sulphonating activity of the enzyme. These results constitute the first study on the molecular cloning, expression, and characterization of a zebrafish cytosolic SULT2 ST.

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