title: Cloning and expression of a novel sulfotransferase with unique substrate specificity from Bombyx mori.
abstract: We identified a cDNA encoding a putative cytosolic sulfotransferase (SULT) by searching the expressed sequence tag database of Bombyx mori, and subsequently obtained the full-length cDNA for this gene via rapid amplification of cDNA ends (RACE). We designated this gene bmST1, and showed by sequence analysis that it belongs to a novel SULT family. The tissue specificity of bmST1 mRNA expression was examined in fifth instar larvae by reverse transcriptase-polymerase chain reaction (RT-PCR), and transcripts were detectable in the silk gland, gut, fat body, and Malpighian tube. A recombinant form of bmST1 was then expressed using a gluthathione S-transferase (GST) gene fusion system, and it was purified from Escherichia coli. Purified bmST1 did not exhibit sulfating activity toward SULT substrates such as 4-nitrophenol, vanillin, hydroxysteroids, or monoamines. Surprisingly, however, recombinant bmST1 showed considerable activity toward 4-nitrocatechol and also gallate esters, although the catechins are not sulfated by this enzyme.
authors.affiliation: Kyoritsu University of Pharmacy, Tokyo, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Hattori
authors.forename: Mio
authors.initials: M
authors.lastname: Hirayama
authors.forename: Hiroko
authors.initials: H
authors.lastname: Suzuki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hamamoto
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Sekimizu
authors.forename: Hiro-Omi
authors.initials: HO
authors.lastname: Tamura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D030541:Databases, Genetic; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D015238:Sulfotransferases