The formation of ceramide-1-phosphate during neutrophil phagocytosis and its role in liposome fusion.
Ceramide, a product of agonist-stimulated sphingomyelinase activation, is known to be generated during the phagocytosis of antibody-coated erythrocytes by polymorphonuclear leukocytes. Agonist-stimulated formation of ceramide-1-phosphate is now shown to occur in 32PO4-labeled neutrophils. Ceramide-1-phosphate is formed by a calcium-dependent ceramide kinase, found predominately in the neutrophil plasma membrane. The neutrophil kinase is specific for ceramide because, in contrast to the bacterial diglyceride kinase, ceramide is not phosphorylated under conditions specific for diglyceride phosphorylation. Conversely, 1,2-diacylglycerol does not serve as substrate for the neutrophil ceramide kinase. Ceramide kinase activation occurs in a time-dependent fashion, reaching peak activity 10 min after formyl peptide stimulation and challenge with antibody-coated erythrocytes. The lipid kinase activity is optimal at pH 6.8. Because the formation of the phagolysosome is a critical event in phagocytosis, the effect of ceramide-1-phosphate in promoting the fusion of liposomes was determined. Both the addition of increasing concentrations of sphingomyelinase D and ceramide-1-phosphate promoted liposomal fusion. In summary, ceramide-1-phosphate is formed during phagocytosis through activation of ceramide kinase. Ceramide-1-phosphate may promote phagolysosome formation.
Hinkovska-Galcheva
V T
VT
Department of Pediatrics, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan 48109, USA.
Boxer
L A
LA
Mansfield
P J
PJ
Harsh
D
D
Blackwood
A
A
Shayman
J A
JA
D002118:Calcium / Q000378:metabolism; D002518:Ceramides / Q000096:biosynthesis*; D004789:Enzyme Activation; D006801:Humans; D066298:In Vitro Techniques; D008081:Liposomes / Q000378:metabolism*; D008561:Membrane Fusion; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D010587:Phagocytosis; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism
D002518:Ceramides; D008081:Liposomes; C065576:ceramide 1-phosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C066324:ceramide kinase; D002118:Calcium