Transverse redistribution of phospholipids during human platelet activation: evidence for a vectorial outflux specific to aminophospholipids.
The redistribution kinetics of phospholipids during human platelet activation by calcium ionophore were investigated to determine the specificity of the observed phospholipid outflux [Bassé et al. (1993) Biochemistry 32, 2337]. (1) Two double-labeling experiments were performed with a combination of equal amounts of spin- and fluorescently-labeled phosphatidylserine and phosphatidylcholine. During A23187-induced activation, 50% of the internal phosphatidylserine analogs were rapidly (t1/2 < 1 min) reexposed on the platelet surface while no reciprocal influx of the external phosphatidylcholine analogs was observed. (2) Treatment with chlorpromazine allowed the internalization of 20% of external spin-labeled sphingomyelin or spin-labeled phosphatidylcholine, without either inducing platelet activation or interfering with aminophospholipid translocase activity or with A23187-induced activation (dense granule secretion and vesicle shedding). During A23187-induced activation, none of the previously internalized choline head phospholipids were exposed externally, while spin-labeled phosphatidylserine outward movements were similar irrespective of whether platelets were pretreated or not pretreated with chlorpromazine. Our results demonstrated that during strong platelet activation (1) the PL excess in the internal leaflet, due to the probe addition, is not responsible for their outflux; (2) the rapid aminophospholipid outflux is definitely a vectorial outflux not counterbalanced by a rapid reciprocal influx of choline head phospholipids (i.e., not scrambling); and (3) the vectorial outflux is specific for aminophospholipids since previously internalized sphingomyelin and phosphatidylcholine did not move outward. This suggests that the specific vectorial outflux of aminophospholipids could be catalyzed by a "reverse aminophospholipid translocase" activity.
Gaffet
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P
URA 1856 CNRS, CP 107, Université Montpellier II, France.
Bettache
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N
Bienvenüe
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D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000097:blood; D001792:Blood Platelets / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002746:Chlorpromazine / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005456:Fluorescent Dyes; D005994:Glycerophosphates / Q000097:blood; D005997:Glycerylphosphorylcholine / Q000031:analogs & derivatives / Q000097:blood; D006801:Humans; D007700:Kinetics; D010713:Phosphatidylcholines / Q000097:blood; D010718:Phosphatidylserines / Q000097:blood; D010743:Phospholipids / Q000097:blood*; D015539:Platelet Activation; D012701:Serotonin / Q000097:blood; D013050:Spectrometry, Fluorescence; D013113:Spin Labels; D014316:Tritium
C078632:1-acyl-2-(12-((7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)dodecanoyl)phosphatidylcholine; C088333:1-palmitoyl-2-(4-doxylpentanoyl)-glycerophosphoserine; D005456:Fluorescent Dyes; D005994:Glycerophosphates; C034296:N-(7-nitro-2,1,3-benzoxadiazol-4-yl)phosphatidylserine; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids; D013113:Spin Labels; D014316:Tritium; D012701:Serotonin; D000001:Calcimycin; D005997:Glycerylphosphorylcholine; C088332:1-palmitoyl-2-(4-doxylpentanoyl)glycerophosphocholine; D009327:4-Chloro-7-nitrobenzofurazan; D002746:Chlorpromazine