Assay of Cellular Diacylglycerol and Monoacylglycerol Lipases
Arachidonic acid has been implicated as a second messenger in numerous systems, and many agonists stimulate the intracellular accumulation of arachidonic acid (1,2). Arachidonic acid may be generated by phospholipase A2 hydrolysis of phospholipids, or by hydrolysis of diacylglycerol through the sequential action of diacylglycerol lipase and monoacylglycerol lipase (3, 4, 5, 6). These activities can be easily measured using sensitive radiometric assays as described herein.
KeywordsVortex Hydrolysis Chloroform Toluene Lipase
- 2.Graber, R., Sumida, C., and Nunez, E. A. (1994) Fatty acids and cell signal trans-duction. J. Lipid Mediat. 9, 91–116.Google Scholar
- 4.Balsinde, J., Diez, E., and Mollinedo, F. (1991) Arachidonic acid release from diacylglycerol in human neutrophils. Translocation of diacylglycerol-deacylating enzyme activities from an intracellular pool to plasma membrane upon cell activation. J. Biol. Chem. 266, 15,638–15,643.PubMedGoogle Scholar