title: Highly conserved mouse and human brain sulfotransferases: molecular cloning, expression, and functional characterization.
abstract: By employing reverse transcription-polymerase chain reaction (RT-PCR) in conjunction with 5'-rapid amplification of cDNA ends technique, we have cloned a novel mouse sulfotransferase cDNA. Database search led to the identification of a human gene encoding the homologue of this newly discovered mouse sulfotransferase. RT-PCR technique was employed to clone the cDNA encoding the human enzyme. Sequence analysis revealed that the novel mouse and human sulfotransferases display nearly 98% identity in their amino acid sequences. Their amino acid sequence identity to other known cytosolic sulfotransferases, however, was found to be below 36%. These two highly conserved sulfotransferases therefore appear to belong to a family different from the two major mammalian cytosolic sulfotransferase gene families. Northern blot analysis revealed the neuronal tissue-specific expression of these two novel sulfotransferases. Recombinant mouse and human brain sulfotransferases, expressed using the pGEX-2TK prokaryotic expression system and purified from transformed Escherichia coli cells, migrated as 33 kD proteins upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified mouse and human brain sulfotransferases displayed enzymatic activities toward endogenous and xenobiotic compounds, including L-triiodothyronine, thyroxine, estrone, p-nitrophenol, 2-naphthylamine, and 2-naphthol. Using mouse brain filtrate as substrate, both brain sulfotransferases were shown to catalyze specifically the sulfation of only a few compounds.
authors.affiliation: Biomedical Research Center, The University of Texas Health Center, 11937 U.S. Highway 271, Tyler, TX 75708, USA.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Sakakibara
authors.forename: Masahito
authors.initials: M
authors.lastname: Suiko
authors.forename: T Govind
authors.initials: TG
authors.lastname: Pai
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Nakayama
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Takami
authors.forename: Junko
authors.initials: J
authors.lastname: Katafuchi
authors.forename: Ming Cheh
authors.initials: MC
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015238:Sulfotransferases / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D015238:Sulfotransferases