Electrospray ionization/mass spectrometric analyses of human promonocytic U937 cell glycerolipids and evidence that differentiation is associated with membrane lipid composition changes that facilitate phospholipase A2 activation

Fong Fu Hsu, Zhongmin Ma, Mary Wohltmann, Alan Bohrer, William Nowatzke, Sasanka Ramanadham, John Turk

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49 Scopus citations

Abstract

Upon differentiation, U937 promonocytic cells gain the ability to release a large fraction of arachidonate esterified in phospholipids when stimulated, but the mechanism is unclear. U937 cells express group IV phospholipase A2 (cPLA2), but neither its level nor its phosphorylation state increases upon differentiation. A group VI PLA2 (iPLA2) that is sensitive to a bromoenol lactone inhibitor catalyzes arachidonate hydrolysis from phospholipids in some cells and facilitates arachidonate incorporation into glycerophosphocholine (GPC) lipids in others, but it is not known whether U937 cells express iPLA2. We confirm that ionophore A23187 induces substantial [3H]arachidonate release from differentiated but not control U937 cells, and electrospray ionization mass spectrometric (ESI/MS) analyses indicate that differentiated cells contain a higher proportion of arachidonate-containing GPC species than control cells. U937 cells express iPLA2 mRNA and activity, but iPLA2 inhibition impairs neither [3H]arachidonate incorporation into nor release from U937 cells. Experiments with phosphatidate phosphohydrolase (PAPH) and phospholipase D (PLD) inhibitors coupled with ESI/MS analyses of PLD-PAPH products indicate that differentiated cells gain the ability to produce diacylglycerol (DAG) via PLD-PAPH. DAG promotes arachidonate release by a mechanism that does not require DAG hydrolysis, is largely independent of protein kinase C, and requires cPLA2 activity. This may reflect DAG effects on cPLA2 substrate state.

Original languageEnglish
Pages (from-to)16579-16589
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number22
DOIs
StatePublished - Jun 2 2000

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