Submitted on September 13, 2007
Revised on December 13, 2007
Accepted on December 20, 2007
Identification and characterization by electrospray mass spectrometry of endogenous Drosophila sphingadienes
Henrik Fyrst, Xinyi Zhang, Deron R. Herr, Hoe Sup Byun, Robert Bittman, Van H. Phan, Greg L. Harris, and Julie D. Saba
Research, Children's Hospital Oakland, Oakland, CA 94609
Corresponding Author: jsaba{at}chori.org
Sphingolipids comprise a complex group of lipids concentrated in membrane rafts and whose metabolites function as signaling molecules. Sphingolipids are conserved in Drosophila, where their tight regulation is required for proper development and tissue integrity. In this study, we identified a new family of Drosophila sphingolipids containing two double bonds in the long chain base. The lipids were found at low levels in wild type flies and accumulated markedly in Drosophila Sply mutants, which do not express sphingosine-1-phosphate lyase and are defective in sphingolipid catabolism. To determine the identity of the unknown lipids, purified whole fly lipid extracts were separated on a C18-HPLC column and analyzed using electrospray mass spectrometry. The lipids contain a long chain base of either 14 or 16 carbons with conjugated double bonds at C4,6. The 4,6-sphingadienes were found as free long chain bases, phosphorylated long chain bases and as the sphingoid base in ceramides. The temporal and spatial accumulation of 4,6-sphingadienes in Sply mutants suggest that these lipids may contribute to muscle degeneration observed in these flies.
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