Platelet agonists enhance the import of phosphatidylethanolamine into human platelets.
It is unknown whether the endocytosis-independent transfer of phospholipids from lipoproteins to platelets is regulated by platelet agonists such as thrombin. The movements of the choline phospholipids phosphatidylcholine and sphingomyelin (labeled with either 14C or the fluorescent pyrenedecanoic acid) between low density lipoproteins and platelets were unaffected by thrombin (0.5 unit/ml). In contrast, thrombin accelerated the import of diacyl phosphatidylethanolamine (PE) and alkenylacyl phosphatidylethanolamine into platelets by about 4-fold. Similarly, thrombin receptor-activating peptide (15 microM), collagen (10 microgram/ml), and ADP (10 microM) enhanced PE uptake. High density lipoprotein particles and egg phosphatidylcholine vesicles were also donors for stimulation of platelet PE import. Part of the [14C]arachidonic acid-labeled PE transferred from low density lipoprotein to platelets activated by thrombin and collagen was metabolized to 14C-eicosanoids. Inhibitors of protein kinase C partially prevented thrombin-induced [14C]PE uptake, while direct activators of protein kinase C increased incorporation of [14C]PE into platelets. Proteinaceous factor(s) recovered in the extracellular medium from ADP- and thrombin-activated platelet suspensions were found to accelerate the transfer of pyrenedecanoic acid-labeled PE between donor and acceptor lipid vesicles. The stimulation of import of ethanolamine phospholipids led to a 2-fold enhancement of the prothrombinase activity of thrombin-activated platelets. Our study demonstrates that physiological platelet stimuli increase specifically the transfer of ethanolamine phospholipids from lipoproteins to platelets through a secretion-dependent mechanism. This might contribute to the increase of procoagulant activity of stimulated platelets.
Physiologisches Institut der Universität München, Pettenkoferstrasse 12, D-80336 München, Germany. bernd.engelmann@med.unimuenchen.de
B
B
Engelmann
B
B
Schaipp
P
P
Dobner
M
M
Stoeckelhuber
C
C
Kögl
W
W
Siess
A
A
Hermetter
D000244:Adenosine Diphosphate / Q000494:pharmacology; D001055:Apolipoproteins B / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism; D001120:Arginine / Q000031:analogs & derivatives; D001692:Biological Transport; D001777:Blood Coagulation; D001792:Blood Platelets / Q000187:drug effects*; D003094:Collagen / Q000494:pharmacology; D003619:Dansyl Compounds / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology; D010714:Phosphatidylethanolamines / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D015539:Platelet Activation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors
D001055:Apolipoproteins B; D003619:Dansyl Compounds; D006629:Hirudins; D008077:Lipoproteins, LDL; D010446:Peptide Fragments; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C076901:thrombin receptor-activating peptide (P508-530); D016718:Arachidonic Acid; D000244:Adenosine Diphosphate; D003094:Collagen; D001120:Arginine; D011493:Protein Kinase C