Sulfation of indoxyl by human and rat aryl (phenol) sulfotransferases to form indoxyl sulfate


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BANOĞLU E. , King R.

EUROPEAN JOURNAL OF DRUG METABOLISM AND PHARMACOKINETICS, cilt.27, sa.2, ss.135-140, 2002 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 27 Konu: 2
  • Basım Tarihi: 2002
  • Doi Numarası: 10.1007/bf03190428
  • Dergi Adı: EUROPEAN JOURNAL OF DRUG METABOLISM AND PHARMACOKINETICS
  • Sayfa Sayıları: ss.135-140

Özet

The aim of this study was to identify sulfotransferase (SULT) isoform(s) responsible for the formation of indoxyl sulfate from indoxyl (3-hydroxyindole). Indoxyl was incubated together with the co-substrate 3'-phosphoadenosine 5'-phosphosulfate (PAPS) and either human or rat liver cytosol or recombinant sulfotransferase enzymes. Formation of indoxyl sulfate from indoxyl was measured by HPLC and used for determination of sulfonation rates. Both cytosols sulfonated indoxyl with apparent K-m values of 6.8 +/- 0.9 muM for human and 3.2 +/- 0.6 muM for rat cytosol. To help identify the isoform(s) of SULT responsible for indoxyl sulfate formation, indoxyl was incubated with human and rat liver cytosols and PAPS in the presence of isoform-specific SULT inhibitors. No inhibition was observed by DHEA, a specific hydroxysteroid sulfotransferase inhibitor, nor by oestrone, an inhibitor of oestrogen sulfotransferase. However, an aryl (phenol) sulfotransferase inhibitor, 2,6-dichloro-4-nitrophenol (DCNP), inhibited the formation of indoxyl sulfate with a IC50 values of 3.2 muM for human and 1.0 muM for rat cytosol indicating that human and rat aryl (phenol) sulfotransferases are responsible for the formation of indoxyl sulfate. When indoxyl was incubated with SULT1A1*2, a human recombinant aryl SULT, an apparent K-m value of 5.6 +/- 1.8 muM was obtained. Kinetic studies with human and rat cytosols and human recombinant SULT1A1*2 gave similar kinetic values indicating that human and rat aryl sulfotransferases efficiently catalyze the formation of indoxyl sulfate, an important uremic toxin metabolite.