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J. Biol. Chem., Vol. 277, Issue 42, 39368-39378, October 18, 2002
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From the Unité INSERM 352, Laboratoire de Biochimie et
Pharmacologie, INSA de Lyon, 69621 Villeurbanne, France,
Docosahexaenoic acid (DHA), an
n-3 polyunsaturated fatty acid that inhibits T lymphocyte
activation, has been shown to stimulate phospholipase D (PLD) activity
in stimulated human peripheral blood mononuclear cells (PBMC). To
elucidate the mechanisms underlying the DHA-induced PLD activation, we
first characterized the PLD expression pattern of PBMC. We show that
these cells express PLD1 and PLD2 at the protein and mRNA level and
are devoid of oleate-dependent PLD activity. DHA enrichment
of PBMC increased the DHA content of cell phospholipids, which was
directly correlated with the extent of PLD activation. The DHA-induced
PLD activation was independent of conventional protein kinase C but
inhibited by brefeldin A, which suggests ADP-ribosylation factor
(ARF)-dependent mechanism. Furthermore, DHA enrichment
dose-dependently stimulated ARF translocation to cell
membranes. Whereas 50% of the guanosine
5'-3-O-(thio)triphosphate plus ARF-dependent
PLD activity and a substantial part of PLD1 protein were located to the
detergent-insoluble membranes, so-called rafts, of non-enriched PBMC,
DHA treatment strongly displaced them toward detergent-soluble
membranes where ARF is present. Collectively, these results suggest
that the exclusion of PLD1 from lipid rafts, due to their partial
disorganization by DHA, and its relocalization in the vicinity of ARF,
is responsible for its activation. This PLD activation might be
responsible for the immunosuppressive effect of DHA because it is known
to transmit antiproliferative signals in lymphoid cells.
The Mechanism of Docosahexaenoic Acid-induced Phospholipase D
Activation in Human Lymphocytes Involves Exclusion of the Enzyme from
Lipid Rafts*
,, Dipartimento di Sanità Pubblica e Biologia
Cellulare, Cattedra di Anatomia Umana, Università di Roma Tor
Vergata, 00173 Rome, Italie, and § Centre de Recherche du
CHUL, Laval University, Ste-Foy,
Québec G1V 4G2, Canada
*
This work was supported by a CNR-INSERM joint program grant
(to C. S. and A. F. P.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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