Human liver dehydroepiandrosterone sulfotransferase: molecular cloning and expression of cDNA.

摘要: Sulfation is an important pathway in the metabolism of many hormones and drugs. Human liver contains at least three well characterized sulfotransferase (ST) enzymes, i.e., dehydroepiandrosterone (DHEA) ST two forms phenol (PST). Our goal was to purify, obtain partial amino acid sequence for, clone express cDNA for human DHEA ST. Polymerase chain reaction primers were designed on basis homology among rat hydroxysteroid ST, PST, bovine estrogen These amplified a unique from cDNA, this polymerase product used screen library. Two clones isolated that contained identical open reading frames, 855 nucleotides, encoded protein 285 acids. The deduced included separate 27- 23-amino sequences those obtained by microsequencing proteolytic fragments purified Translation, rabbit reticulocyte lysate system, mRNA transcribed vitro resulted 35-kDa translation comigrated with during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This also catalyzed sulfation but not model substrates PST found liver. create expression constructs eukaryotic vector P91023(B), these transfect COS-1 cells. transfected cells expressed high level activity, enzyme activity displayed pattern inhibition inhibitor 2,6-dichloro-4-nitrophenol Cloning sulfate-conjugating will enhance understanding relationship between other STs, as enzymes species.