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J. Biol. Chem., Vol. 268, Issue 2, 839-844, Jan, 1993
M Murakami, I Kudo and K Inoue
A lysate of unstimulated human umbilical vein endothelial cells (HUVEC)
exhibited phospholipase A2 (PLA2) activity, which hydrolyzed phospholipids
bearing arachidonate more preferentially than those bearing linoleate at
the sn-2 position. An anti-rabbit cytosolic PLA2 monoclonal antibody
absorbed the activity, whereas an anti-human type II PLA2 monoclonal
antibody did not. HUVEC treated with thrombin generated prostaglandin I2
(PGI2), and the PLA2 activity of the thrombin-stimulated cells was absorbed
almost completely by the anti- cytosolic PLA2 antibody. HUVEC treated with
tumor necrosis factor (TNF) also generated PGI2. PGI2 generation by
TNF-treated cells was suppressed partially by extracellular addition of the
anti-type II PLA2 antibody. PLA2 activity in a lysate of TNF-stimulated
cells was increased about 2-3-fold, and about half of the increased
activity was suppressed by the anti-type II PLA2 antibody. Addition of
heparin together with TNF resulted in release of type II PLA2 in the
medium. Thus, both cytosolic and type II PLA2s may be involved in agonist-
stimulated PGI2 synthesis in HUVEC. Furthermore, exogenously added type II
PLA2 was bound to the cell surface and synergistically enhanced PGI2
generation in TNF-stimulated HUVEC. This binding was blocked by either
heparin or a monoclonal antibody recognizing the heparin-binding domain of
type II PLA2. Taken together, type II PLA2 generated endogenously as well
as added exogenously may be captured on the HUVEC surface via heparan
sulfate proteoglycan and may contribute to cellular arachidonate
metabolism.
Molecular nature of phospholipases A2 involved in prostaglandin I2 synthesis in human umbilical vein endothelial cells. Possible participation of cytosolic and extracellular type II phospholipases A2
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
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P. Ancian, Gér. Lambeau, M.-G.èv. Mattéi, and M. Lazdunski The Human 180-kDa Receptor for Secretory Phospholipases A[IMAGE] J. Biol. Chem., April 14, 1995; 270(15): 8963 - 8970. [Abstract] [Full Text] [PDF]
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