Journal
JOURNAL OF LIPID RESEARCH
Volume 50, Issue 10, Pages 1986-1995Publisher
ELSEVIER
DOI: 10.1194/jlr.M800367-JLR200
Keywords
ceramide-1-phosphate; ceramide kinase; prostaglandins; phospholipase A(2); inflammation; arachidonic acid; dodecane; eicosanoids
Categories
Funding
- Veteran's Administration
- National Institutes of Health [HL072925, CA117950, 1C06-RR17393]
- America Heart Association [AHA 5-30693, AHA 0625502U]
- Ministerio de Education y Ciencia-Spain [BFU2006-13689]
- Virginia Commonwealth University Dept. of Neurobiology & Anatomy Microscopy Facility
- NIH-NINDS Center core [5P30NS047463]
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Previously, our laboratory demonstrated that ceramide-1-phosphate (C1P) specifically activated group IVA cytosolic phospholipase A(2) (cPLA(2)alpha) in vitro. In this study, we investigated the chain length specificity of this interaction. C1P with an acyl-chain of >= 6 carbons efficiently activated cPLA(2)alpha in vitro, whereas C-2-C1P, was unable to do so. Delivery of C1P to cells via the newly characterized ethanol/dodecane system demonstrated a lipid-specific activation of cPLA(2)alpha, AA release, and PGE(2) synthesis (EC50 = 400 nM) when compared to structurally similar lipids. C1P delivered as vesicles in water also induced a lipid-specific increase in AA release. Mass spectrometric analysis demonstrated that C1P delivered via ethanol/dodecane induced a 3-fold increase in endogenous C1P with little metabolism to ceramide. C1P was also more efficiently delivered (>3-fold) to internal membranes by ethanol/dodecane as compared to vesiculated C1P. Using this now established delivery method for lipids, C-2-C1P was shown to be ineffective in the induction of AA release as compared with C-6-C1P, C-16-C1P, and C-18:1 C1P. Here, we demonstrate that C1P requires = 6 carbon acyl-chain to activate cPLA(2)alpha. Thus, published reports on the biological activity of C-2-C1P are not via eicosanoid synthesis. Furthermore, this study demonstrates that the alcohol/dodecane system can be used to efficiently de-liver exogenous phospholipids to cells for the examination of specific biological effects.-Wijesinghe, D. S., P. Subramanian, N. F. Lamour, L. B. Gentile, M. H. Granado, A. Bielawska, Z. Szulc, A. Gomez-Munoz, and C. E. Chalfant. Chain length specificity for activation of cPLA(2)alpha by C1P: use of the dodecane delivery system to determine lipid-specific effects. J. Lipid Res. 2009. 50: 1986-1995.
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