Does PAPS generation determine the overall sulfate conjugation in human platelets?

The formation of 3'-phospho-adenosine-5'phosphosulfate (PAPS) depends on essentially two enzymic reactions catalyzed by ATP-sulfurylase and APS-kinase (adenosine 5'-phosphosulfate-kinase). In this paper, PAPS generation by human platelets was determined by the transfer of 35sulfate from PAP 35S formed in vitro to N-acetyldopamine (NADA), using the phenolsulfotransferase extracted from rat liver. A pre-requisite of this quantitative procedure was the prior inhibition of the sulfate-activating system in the latter enzyme preparation. This was accomplished by the addition of 10 mM EDTA and 14 mM pyrophosphate. The PAPS-generating system of human platelets exhibited two pH peaks with higher activity at pH 8 than pH 6. Optimal concentrations of ATP and Mg++ at 7 mM were required for the two reactions. PAPS generation so measured showed a highly significant correlation with the overall sulfate conjugation of NADA: a correlation coefficient of 0.96 was established from data obtained from 60 platelet preparations of normal subjects