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J. Biol. Chem., Vol. 263, Issue 5, 2217-2222, 02, 1988
ML Nieto, S Velasco and M Sanchez Crespo
The generation of platelet-activating factor (PAF) in response to
complement-coated zymosan particles, ionophore A23187 and 12-O-
tetradecanoylphorbol 13-acetate (TPA) was studied in human
polymorphonuclear leukocytes (PMN). TPA was an active stimulator of PAF
biosynthesis but showed a time course more protracted than that observed in
response to other secretagogues. TPA was not found to activate
acetyl-CoA:lyso-PAF acetyltransferase activity nor to significantly enhance
the incorporation of either [3H] lyso-PAF or [3H]acetate into a lipid
fraction co-migrating as PAF. To assess whether TPA could be a secretagogue
that promotes the biosynthesis of PAF through the
dithiothreitol-insensitive cholinephosphotransferase pathway, this enzymic
activity was assayed in homogenates from PMN preincubated in the presence
of several secretagogues. None of the agonists was found to enhance this
enzyme activity. However, TPA did enhance the incorporation of
[methyl-3H]choline and both alkyl- and acetyl-labeled
1-O-hexadecyl-2-acetyl-sn-glycerol into a lipid fraction co-migrating with
PAF. This incorporation showed a time course parallel to that of the
generation of PAF in response to TPA. Incubation of
[methyl-3H]choline-labeled cells in the presence of 1-O-hexadecyl-2-
acetyl-sn-glycerol caused an enhanced incorporation of the label into the
fraction co-migrating as PAF, and this incorporation was synergistically
enhanced by TPA. Pulse-chase experiments with choline showed that TPA
caused an early accumulation of choline into phosphatidylcholine and PAF
rather than into CDP-choline. The present data indicate that TPA is the
only agonist that could initiate the biosynthesis of PAF in human PMN
through the cholinephosphotransferase pathway and that this process of
biosynthesis is regulated at an enzyme step other than
dithiothreitol-insensitive cholinephosphotransferase.
Biosynthesis of platelet-activating factor in human polymorphonuclear leukocytes. Involvement of the cholinephosphotransferase pathway in response to the phorbol esters
Instituto de Investigaciones Medicas de la Fundacion Jimenez Diaz, Centro Asociado al Consejo Superior de Investigaciones Cientificas, Madrid, Spain.
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