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Journal of Biochemistry Advance Access originally published online on June 8, 2009
Journal of Biochemistry 2009 146(3):399-405; doi:10.1093/jb/mvp087
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© The Authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved

Molecular Cloning, Expression and Characterization of A Novel Mouse SULT6 Cytosolic Sulfotransferase

Saki Takahashi1,2, Yoichi Sakakibara1,2,*, Emi Mishiro1,2, Haruna Kouriki1, Rika Nobe1,2, Katsuhisa Kurogi1, Shin Yasuda3,4, Ming-Cheh Liu3 and Masahito Suiko1,2

1Department of Biochemistry and Applied Biosciences, University of Miyazaki, Miyazaki, Miyazaki 889-2192; 2United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Kagoshima 890-0065, Japan; 3Department of Pharmacology, College of Pharmacy, The University of Toledo, Toledo, OH 43606, USA; and 4Department of Bioscience, School of Agriculture, Tokai University, Aso, Kumamoto 869-1404, Japan

*To whom correspondence should be addressed. Tel: +81-985-58-7211, Fax: +81-985-58-7211, E-mail: ysakaki{at}cc.miyazaki-u.ac.jp

Received March 31, 2009; Accepted May 19, 2009


  Abstract

By searching the mouse EST database, we identified a novel mouse cytosolic sulfotransferase (SULT) cDNA (RIKEN cDNA 2410078J06). Sequence analysis revealed that this new SULT belongs to the cytosolic SULT6 gene family. The recombinant form of this newly identified SULT, designated SULT6B1, was expressed using the pGEX-4T-1 glutathione S-transferase fusion system and purified from transformed BL21 Escherichia coli cells. Purified mouse SULT6B1 exhibited sulfonating activity toward thyroxine and bithionol among a variety of endogenous and xenobiotic compounds tested as substrates. pH optimum of purified mouse SULT6B1 was determined to be 8.0. Tissue-specific expression of mouse and human SULT6B1 was examined by RT–PCR. While human SULT6B1 was specifically expressed in kidney and testis, mouse SULT6B1 was detected in brain, heart, kidney, thymus, lung, liver and testis. Further studies are needed in order to clarify the role of SULT6B1 in the metabolism of thyroxine and possibly some xenobiotics in mouse.

Key Words: detoxification, metabolism, sulfonation, sulfotransferase, thyroxine

Abbreviations: 5'-PSB motif, 5'-phosphosulfate binding motif; 3'-PB motif, 3'-phosphate binding motif; PAPS, 3'-phosphoadenosine 5'-phosphosulfate; RT–PCR, reverse transcription–polymerase chain reaction; SDS–PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; SULT, sulfotransferase


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