Acetyltransferase activity and production of platelet-activating factor by human neutrophils activated with various...

The acetyltransferase activity and biosynthesis of platelet-activating factor (PAF) were assessed in human neutrophils activated by 4 microM A23187, 20 ng/ml phorbol myristate acetate (PMA), and 10(-6) M n-formyl-methionyl-leucyl-phenylalanine (fMLP). All three agents elevated the acetyltransferase activity dose-dependently. There were no significant differences in the Km values for acetyl CoA between non-stimulated and stimulated cells. All three stimuli gave a similar Vmax value of acetyltransferase determined 10 min after stimulation, being more than twice as high as the control value. By contrast, the amount of PAF produced by the neutrophils differed with the stimuli, A23187 being by far the most potent. The time course of PAF synthesis, particularly when activated by PMA, did not parallel that of acetyltransferase activity; PMA-induced PAF production was negligible for the first 20 min and gradually increased to reach its plateau 60 min after stimulation, while enzyme activity was at its highest level 5 min after stimulation. These results suggest that the different stimuli activated the same acetyltransferase, and that there was an increase in the number of the enzyme molecules in the activated state. It is unlikely that a change in the biological activity of a preexisting enzyme without a change in the number of active enzyme molecules could be the cause. Certain factors other than acetyltransferase may also regulate the PAF biosynthesis induced by these stimuli.