The different effects of sphingosine on diacylglycerol kinase isozymes in Jurkat cells, a human T-cell line.
We studied the effect of sphingosine on the activities of soluble and membrane-bound isozymes from Jurkat cells using combinations of different substrates (arachidonoyl- and didecanoyl DGs) and assay methods (octylglucoside mixed micellar and deoxycholate suspension assays). The results suggested the presence of at least four DGK isoforms, which could be distinguished from each other with respect to intracellular localization, specificity to DG molecular species, responsiveness to sphingosine, and reactivity to anti-80 kDa DGK antibody. We confirmed the presence of arachidonoyl DG-specific DGK in membranes, though this isozyme was not activated by sphingosine. We detected in the cytosol at least two species of sphingosine-activatable and non-activatable DGK isoforms, the major species being the 80 kDa DGK. We postulate that both or either of the two soluble DGKs may be the target of the sphingosine action.
Yamada
K
K
Department of Biochemistry, School of Medicine, Sapporo Medical University, Japan.
Sakane
F
F
D002460:Cell Line / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D003840:Deoxycholic Acid; D019852:Diacylglycerol Kinase; D004075:Diglycerides / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005960:Glucosides; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D013110:Sphingosine / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000201:enzymology
D004075:Diglycerides; D005960:Glucosides; D007527:Isoenzymes; D003840:Deoxycholic Acid; C018619:octyl-beta-D-glucoside; D017853:Phosphotransferases (Alcohol Group Acceptor); D019852:Diacylglycerol Kinase; D013110:Sphingosine