Molecular Cloning and Characterization of a Novel Canine Sulfotransferase
Sulfonation, as mediated by the cytosolic sulfotransferases (SULTs), represents an important mechanism in vivo for the detoxification/metabolism of drugs and other xenobiotics, as well as endogenous neurotransmitters and hormones. Madin-Darby canine kidney (MDCK) cells have been widely used as a model system for investigating the excretion/detoxification mechanism of kidney. In this study, a cDNA encoding a novel canine SULT was cloned from MDCK cells using reverse transcription-polymerase chain reaction (RT-PCR) technique. Nucleotide sequence revealed that the newly cloned canine SULT cDNA has an open reading frame encompassing 912 nucleotides and encoding a 303-amino acid polypeptide. Analysis of the deduced amino acid sequence showed that the novel canine SULT is highly homologous to human SULT1C4, with 82.8% amino acid identity. Recombinant canine SULT1C4, expressed and purified using the pGEX-4T-1 GST gene fusion system, exhibited strong phenol-sulfonating activities toward 1- or 2-naphthol and o-bromophenol among a variety of xenobiotic compounds tested as substrates. These data indicated that the newly identified canine SULT1C4 may function in kidney for the detoxification of xenobiotic compounds.
KeywordsMDCK Cell Amino Acid Sequence Identity Xenobiotic Compound Canine Tissue Phenol Sulfotransferases
This work was supported by a Grant-in-Aid for Scientific Research (B), (C) (M.S. and Y.S.), a Grant-in Aid for Encouragement of Young Scientists (Y.S.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, Health and Sciences Research Grants (Toxicogenomics) from the Ministry of Health, Labor and Welfare of Japan (Y.S.) and start-up funds from College of Pharmacy, The University of Toledo (M.C.L.). We also thank Satoshi Miyazaki, Saki Takahashi, Dr Madhyasha Harish Kumar for great help of this study and manuscript.
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