Stimulation of cellular sphingomyelin import by the chemokine connective tissue-activating peptide III.
The selective import of phospholipids into cells could be mediated by proteins secreted from the cells into the extracellular compartment. We observed that the supernatants obtained from suspensions of thrombin-activated platelets stimulated the exchange of pyrene (py)-labeled sphingomyelin between lipid vesicles in vitro. The proteins with sphingomyelin transfer activity were purified and identified as the chemokine connective tissue-activating peptide III (CTAP-III) and platelet basic protein. Isolated CTAP-III stimulated the exchange of py-sphingomyelin between lipid vesicles but did not affect the translocations of py-labeled phosphatidylcholine and phosphatidylethanolamine. CTAP-III rapidly increased the transfer of py-sphingomyelin from low density lipoproteins into peripheral blood lymphocytes, other immune cells, and fibroblasts. In the presence of heparin, CTAP-III was unable to insert sphingomyelin into the peripheral blood lymphocytes. The activation energy of the py-sphingomyelin transfer suggested that the translocation proceeded entirely in a hydrophobic environment. [(3)H]Sphingomyelin transferred to the cells by CTAP-III was hydrolyzed to [(3)H]ceramide and [(3)H]sphingosine after activation with tumor necrosis factor alpha. The generation of the [(3)H]sphingolipid messengers was catalyzed by acid sphingomyelinase. Our results identify CTAP-III as the first mediator of the selective (endocytosis-independent) cellular import of sphingomyelin allowing the paracrine modulation of the sphingolipid signaling.
Stoeckelhuber
M
M
Physiologisches Institut der Universität München, Schillerstrasse 44, 80336 München, Germany.
Dobner
P
P
Baumgartner
P
P
Ehlert
J
J
Brandt
E
E
Mentele
R
R
Adam
D
D
Engelmann
B
B
D001692:Biological Transport; D001779:Blood Coagulation Factors / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D010455:Peptides; D013109:Sphingomyelins / Q000378:metabolism*
D001779:Blood Coagulation Factors; D010455:Peptides; D013109:Sphingomyelins; C009862:connective tissue-activating peptide; C018329:low affinity platelet factor 4