Molecular cloning, expression, and enzymatic characterization of rabbit hydroxysteroid sulfotransferase AST-RB2

Journal of Biochemistry
K YoshinariY Yamazoe

Abstract

Cytosolic sulfotransferases, which consist of at least three gene families, play a major role in activation and detoxification of both endogenous and exogenous chemicals. We recently purified a rabbit sulfotransferase, AST-RB2, showing high activities to both hydroxysteroids and amines. To characterize this enzyme, a rabbit cDNA library was screened using anti-AST-RB2 antibodies. The isolated cDNA was judged to encode AST-RB2 (ST2A8) based on the amino acid sequences of peptide fragments obtained from purified AST-RB2. The cDNA showed high similarity to other mammalian hydroxysteroid sulfotransferases (ST2) at the amino acid level (58-68%), but low similarity to aryl sulfotransferases (ST1) (less than 37%). The protein expressed in Escherichia coli catalyzed sulfation of typical ST2 substrates. Therefore, ST2A8 was judged to belong to the ST2 family from both its primary structure and substrate specificity. The ST2A8 protein expressed in E. coli clearly differed from rat ST2A1 and ST2A2 on its localization (cytosol/insoluble fraction ratio). ST2A8 had no activity to lithocholate, but showed the highest catalysis on dehydroepiandrosterone and testosterone among the four forms (ST2A1, ST2A2, ST2A3, and ST2A8), indicating a clear ...Continue Reading

Citations

Sep 30, 1999·Cancer Letters·Y YamazoeK Iwasaki
May 15, 2002·The International Journal of Biochemistry & Cell Biology·E RileyM E McManus
Sep 11, 1999·The International Journal of Biochemistry & Cell Biology·N HehonahM McManus
Jun 3, 2000·Annual Review of Pharmacology and Toxicology·K Nagata, Y Yamazoe
May 29, 2004·Pharmacogenetics·Rebecca L BlanchardMichael W H Coughtrie
May 8, 2009·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Laddawan SenggunpraiYasushi Yamazoe
Sep 26, 2008·The Journal of Pharmacology and Experimental Therapeutics·Laddawan SenggunpraiYasushi Yamazoe

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