Open AccessDiacylglycerol kinase‐dependent formation of phosphatidic acid molecular species during interleukin‐2 activation in CTLL‐2 T‐lymphocytes
Author(s) -
Mizuno Satoru,
Sakai Hiromichi,
Saito Masafumi,
Kado Sayaka,
Sakane Fumio
Publication year - 2012
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1016/j.fob.2012.08.006
Subject(s) - diacylglycerol kinase , phosphatidic acid , phospholipid , chemistry , intracellular , biochemistry , kinase , lipidomics , stimulation , protein kinase c , biology , endocrinology , membrane
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.