title: Diacylglycerol kinase-dependent formation of phosphatidic acid molecular species during interleukin-2 activation in CTLL-2 T-lymphocytes.
abstract: Although effective liquid chromatography (LC)/mass spectrometry (MS) methods enabling the separation of phospholipid molecular species have been developed, there are still problems with an intracellular signaling molecule, phosphatidic acid (PA). In this study, we optimized LC/MS conditions to improve the quantitative detection of PA molecular species from a cellular lipid mixture. Using the newly developed LC/MS method, we showed that stimulation of CTLL-2 murine T-lymphocytes by interleukin-2 (IL-2) induced a significant increase of 36:1-, 36:2-, 40:5- and 40:6-diacyl-PA. A diacylglycerol kinase (DGK) inhibitor, R59949, attenuated the increase of 36:1-, 40:5-, 40:6-diacyl-PA, suggesting that DGK IL-2-dependently and selectively generated these diacyl-PA species.
authors: Department of Chemistry, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.
authors: Satoru
authors: S
authors: Mizuno
authors: Hiromichi
authors: H
authors: Sakai
authors: Masafumi
authors: M
authors: Saito
authors: Sayaka
authors: S
authors: Kado
authors: Fumio
authors: F
authors: Sakane
keywords: DGK, diacylglycerol kinase; DMEM, Dulbecco's modified Eagle's medium; Diacylglycerol kinase; ESI, electrospray ionization; FBS, fetal bovine serum; IL-2, interleukin-2; Interleukin-2; LC, liquid chromatography; Liquid chromatography/electrospray-ionization mass spectrometry; MEF, mouse embryonic fibroblast; MS, mass spectrometry; PA, phosphatidic acid; PLD, phospholipase D.; Phosphatidic acid
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