Purification and kinetic characterization of a dopamine-sulfating form of phenol sulfotransferase from human brain

Biochemistry. 1985 May 7;24(10):2477-82. doi: 10.1021/bi00331a013.


The kinetic and biochemical properties of a purified, monoamine-sulfating form of phenol sulfotransferase (M-PST) from human brain are described. M-PST activity was separated and purified from phenol-sulfating activity by anion-exchange chromatography on DEAE-cellulose and subsequently purified on AffiGel Blue and Sephacryl S-200, routinely giving a final purification of over 20 000-fold, with approximately a 3% yield. The molecular weight of the active species, as estimated by gel filtration chromatography, was 250 000. The purified enzyme was inhibited by NaCl (50% at 325 mM) and showed an optimum for dopamine sulfation at pH 7.0. Of the monoamine substrates examined, 4-methoxytyramine was the most extensively sulfated at 20 microM, while at higher substrate concentrations (200 microM), tyramine was the apparent preferred substrate. Kinetic analysis demonstrated that sulfation by M-PST proceeds via an ordered, bisubstrate reaction mechanism, where 3'-phosphoadenosine 5'-phosphosulfate (PAPS) is the leading substrate. True Km values for dopamine and PAPS were 2.9 and 0.35 microM, respectively. The product inhibitor 3'-phosphoadenosine 5'-phosphate possessed a Ki of 0.07 microM, while the dead-end inhibitor ATP exhibited a Ki of 170 microM.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / pharmacology
  • Arylsulfotransferase
  • Cerebral Cortex / enzymology*
  • Dopamine / analogs & derivatives*
  • Dopamine / analysis
  • Humans
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Weight
  • Osmolar Concentration
  • Substrate Specificity
  • Sulfurtransferases / isolation & purification*
  • Sulfurtransferases / metabolism


  • dopamine 4-O-sulfate
  • dopamine 3-O-sulfate
  • Adenosine Triphosphate
  • Sulfurtransferases
  • Arylsulfotransferase
  • Dopamine