標題: Identification of novel hydroxysteroid-sulfating cytosolic SULTs, SULT2 ST2 and SULT2 ST3, from zebrafish: Cloning, expression, characterization, and developmental expression
作者: Yasuda, Shin
Liu, Ming-Yih
Yang, Yuh-Shyong
Snow, Rhodora
Takahashi, Saki
Liu, Ming-Cheh
生醫工程研究所
Institute of Biomedical Engineering
關鍵字: sulfotransferase;SULT;sulfation;dehydroepiandrosterone;corticosterone;molecular cloning;developmental expression;zebrafish
公開日期: 1-Nov-2006
摘要: By searching the expressed sequence tag database, two zebrafish cDNAs encoding putative cytosolic sulfotransferases (SULTs) were identified. Sequence analysis indicated that these two zebrafish SULTs belong to the cytosolic SULT2 gene family. The recombinant form of these two novel zebratish SULTs, designated SULT2 ST2 and SULT2 ST3, were expressed using the pGEX-2TK glutathione S-transferase (GST) gene fusion system and purified from transformed BL21 (DE3) Escherichia coli cells. Purified GST-fusion protein form of SULT2 ST2 and SULT2 ST3 exhibited strong sulfating activities toward dehydroepiandrosterome (DHEA) and corticosterone, respectively, among various endogenous compounds tested as substrates. Both enzymes displayed pH optima at similar to 6.5. Kinetic constants of the two enzymes, as well as the GST-fusion protein form of the previously identified SULT2 ST1, with DHEA and corticosterone as substrates were determined. Developmental stage-dependent expression experiments revealed distinct patterns of expression of SULT2 ST2 and SULT2 ST3, as well as the previously identified SULT2 ST1, during embryonic development and throughout the larval stage onto maturity. (c) 2006 Elsevier Inc. All rights reserved.
URI: http://dx.doi.org/10.1016/j.abb.2006.09.004
http://hdl.handle.net/11536/11583
ISSN: 0003-9861
DOI: 10.1016/j.abb.2006.09.004
期刊: ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
Volume: 455
Issue: 1
起始頁: 1
結束頁: 9
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