Activation of endothelial cell phospholipase D by sphingosine and sphingosine-1-phosphate.

V. Natarajan, H. N. Jayaram, W. M. Scribner, J. G. Garcia

Research output: Contribution to journalArticle

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Abstract

We have investigated the activation of phospholipase D (PLD) by sphingosine and its derivatives in bovine pulmonary artery endothelial cells (BPAEC) prelabeled with [32P]orthophosphate or [32P]lyso phospholipids. Sphingosine, in a dose- and time-dependent manner, stimulated the hydrolysis of [32P]phosphatidylcholine (PC) resulting in the production of [32P]phosphatidic acid (PA), suggesting PLD activation. In the presence of ethanol (150 mM), the accumulation of [32P]phosphatidylethanol was also observed. The sphingosine-induced stimulation of PLD activity was not affected by treatment with the protein kinase C (PKC) inhibitor staurosporine or by down-regulation of PKC with TPA and was independent of extracellular Ca2+, suggesting that the PLD activation was independent of PKC and Ca2+. Chelation of intracellular Ca2+ with BAPTA actually potentiated the sphingosine-stimulated [32P]PC hydrolysis. Furthermore, the activation of PLD by sphingosine was not abolished by treatment of BPAEC with either cholera or pertussis toxin, indicating noninvolvement of toxin-sensitive G-proteins. In addition to hydrolysis of [32P]PC, sphingosine also stimulated PLD-mediated hydrolysis of [32P]phosphatidylethanolamine and [32P]phosphatidylinositol. Among the various sphingoid compounds, in addition to sphingosine, only sphingosine-1-phosphate (Sph-1-P) activated the endothelial cell PLD. The effect of sphingosine and Sph-1-P on PA phosphatase (PA Pase) activity was tested using [3H]glycerol-labeled PA. The Mg(2+)-independent and membrane-associated PA Pase activity was inhibited by sphingosine (IC50 = 200 microM) but not by Sph-1-P. This implies that sphingosine and Sph-1-P share a similar PLD-stimulating property but differ in their PA Pase inhibitory activity.

Original languageEnglish
Pages (from-to)221-229
Number of pages9
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume11
Issue number2
StatePublished - Aug 1994

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Phospholipase D
Sphingosine
Endothelial cells
Endothelial Cells
Chemical activation
Phosphatidic Acids
Hydrolysis
Phosphatidylcholines
Protein Kinase C
Pulmonary Artery
Phosphatidate Phosphatase
sphingosine 1-phosphate
Staurosporine
Protein C Inhibitor
Cholera Toxin
Pertussis Toxin
Protein Kinase Inhibitors
Chelation
Phosphatidylinositols
GTP-Binding Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Activation of endothelial cell phospholipase D by sphingosine and sphingosine-1-phosphate. / Natarajan, V.; Jayaram, H. N.; Scribner, W. M.; Garcia, J. G.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 11, No. 2, 08.1994, p. 221-229.

Research output: Contribution to journalArticle

Natarajan, V. ; Jayaram, H. N. ; Scribner, W. M. ; Garcia, J. G. / Activation of endothelial cell phospholipase D by sphingosine and sphingosine-1-phosphate. In: American Journal of Respiratory Cell and Molecular Biology. 1994 ; Vol. 11, No. 2. pp. 221-229.
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