Purification and characterization of two amine N-sulfotransferases, AST- RB1 (ST3A1) and AST-RB2 (ST2A8), from liver cytosols of male rabbits

Toshifumi Shiraga, Kazuhide Iwasaki, Takehisa Hata, Kouichi Yoshinari, Kiyoshi Nagata, Yasushi Yamazoe, Yasuo Ohno

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6 Citations (Scopus)

Abstract

Two sulfotransferases (STs), designated as AST-RB1 (ST3A1) and AST-RB2 (ST2A8), with high a amine N-sulfonating activity, were purified from male rabbit liver cytosols. AST-RB1 and AST-RB2 were purified to homogeneity by the anion-exchange, affinity, and hydroxyapatite chromatography. The N- terminus of both enzymes were blocked. The subunit molecular mass of both enzymes was estimated to be 34 kDa on SDS-PAGE. AST-RB1 efficiently catalyzed N-sulfonation of alicyclic, alkyl, and arylamines such as 4-phenyl-1,2,3,6- tetrahydropyridine, 1-[(5-chloro-2-oxo-3(2H)-benzothiazolyl)acetyl]- piperazine, desipramine, and aniline, whereas its catalytic activities toward 2-naphthol and dehydroepiandrosterone (DHEA) were very low. On the other hand, AST-RB2 efficiently catalyzed sulfonation of desipramine and DHEA, but had no activity toward 2-naphthol. Amino acid sequences of peptide fragments derived from the purified AST-RB1 showed no significant homology with previously reported STs, but those from the purified AST-RB2 shared a high similarity with those of the ST2 family. Both enzymes were expressed specifically in the liver. The present results strongly suggest that the purified AST-RB1 is a novel enzyme in terms of structure and catalytic properties showing high selectivity for amine substrates, and AST-RB2 is a quite unique from among ST2A enzymes of other species in its substrate specificity.

Original languageEnglish
Pages (from-to)265-274
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume362
Issue number2
DOIs
Publication statusPublished - 1999 Feb 15

Keywords

  • Cytosol
  • Liver
  • Purification
  • Rabbit
  • Sulfotransferase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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