Acceleration by ceramide of calcium-dependent translocation of phospholipase A2 from cytosol to membranes in platelets.
The effect of ceramide on Ca2+-dependent translocation of cytosolic phospholipase A2 (cPLA2) to membranes was studied. Pretreatment of platelets with sphingomyelinase or C6-ceramide (N-hexanoylsphingosine) led to apparent enhancement of Ca2+-ionophore A23187-stimulated arachidonic acid release but did not affect the cytosolic phospholipase A2 (cPLA2) activity. Under these conditions, the cPLA2 proteins in membranes increased significantly, compared with those by A23187 alone. Sphingomyelinase and C6-ceramide, but not C6-dihydroceramide, a control analog of C6-ceramide, also facilitated the Ca2+-dependent increase in the cPLA2 protein, as well as the activity, in membranes induced by addition of Ca2+ into platelet lysate. Protein kinase Calpha, which possesses a Ca2+-dependent lipid binding domain, was increased in membranes in a Ca2+-dependent manner, but the increase was not accelerated by sphingomyelinase or C6-ceramide. These findings suggest that ceramide in membranes potentiates Ca2+-dependent cPLA2 translocation from cytosol to membranes, probably through modification of membrane phospholipid organization.
Kitatani
K
K
Department of Pathological Biochemistry, Kyoto Pharmaceutical University, Japan.
Oka
T
T
Murata
T
T
Hayama
M
M
Akiba
S
S
Sato
T
T
D000818:Animals; D016718:Arachidonic Acid / Q000097:blood; D001693:Biological Transport, Active / Q000187:drug effects; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism*; D002118:Calcium / Q000097:blood*; D002518:Ceramides / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000097:blood; D008563:Membrane Lipids / Q000097:blood; D008566:Membranes / Q000378:metabolism; D010741:Phospholipases A / Q000097:blood*; D054467:Phospholipases A2; D010743:Phospholipids / Q000097:blood; D011493:Protein Kinase C / Q000097:blood; D051571:Protein Kinase C-alpha; D011817:Rabbits; D013108:Sphingomyelin Phosphodiesterase / Q000494:pharmacology
D002518:Ceramides; D007527:Isoenzymes; D008563:Membrane Lipids; D010743:Phospholipids; C101954:N-caproylsphingosine; D016718:Arachidonic Acid; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D010741:Phospholipases A; D054467:Phospholipases A2; D013108:Sphingomyelin Phosphodiesterase; D002118:Calcium