Phospholipid transverse mobility modifications in plasma membranes of activated platelets: An ESR study

Bassé, François; Gaffet, Patrick; Rendu, Francine; Bienvenüe, Alain

doi:10.1016/0006-291x(92)91581-a

Cited by 10 publications

(8 citation statements)

References 18 publications

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“…Double labeling enabled us (1) to monitor the redistribution kinetics of aminoPLs and choline head PLs at the same time in the same platelet suspension and (2) to suppress the initial imbalance in the plasma membrane due to one or another probe. PC analogs are incorporated entirely in the outer leaflet of the platelet plasma membrane within 2 min and remain so for at least 1.5 h without internalization (NBD-PC) (data not shown) or redistributed slowly (tu2 > 20 min) to the inner leaflet [(0,2)PC] (Basse et al, 1992). PS analogs are incorporated in the outer leaflet of the plasma membrane within 2 min (data not shown), and their subsequent internalization to the inner leaflet is both rapid (t\n < 5 min) and ATP-dependent (Seigneuret & Devaux, 1984;Zachowski et al, 1986;Sune et al, 1987;Connor et al, 1992).…”

Section: Resultsmentioning

confidence: 99%

“…The amount of hydrolyzed NBD-PL (C6-NBD) in the platelet suspension was expressed as a percentage of total NBD-PL PC]}, and sphingomyelin {/V-(4-doxylpentanoyl)-franssphingenyl-l-phosphocholine [(0,2)SM]} were synthesized as described previously (Davoust et al, 1983). The redistribution of the paramagnetic probes previously incorporated in the outer leaflet of the plasma membrane was quantified as follows (Morrot et al, 1989;Basse et al, 1992). Spinlabeled analogs were added (1-2% of endogenous PL) to the platelet suspension at time zero from a concentrated solution in buffer B. Spin-labeled PLs [(0,2)PLs] were incorporated into the outer plasma membrane leaflet in less than 1 min (Seigneuret & Devaux, 1984).…”

Section: Methodsmentioning

confidence: 99%

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Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Gaffet¹,

Bettache²,

Bienvenüe³

1995

Biochemistry

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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.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Gaffet¹,

Bettache²,

Bienvenüe³

1995

Biochemistry

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“…Spin‐labelled analogues of PS {1‐palmitoyl‐2‐(4‐doxylpentanoyl)‐glycerophosphoserine [(0,2)PS]}, and PC {1‐palmitoyl‐2‐(4‐doxylpentanoyl)‐glycerophosphocholine [(0,2)PC]} were synthesized as described previously ( Davoust et al , 1983 ). The redistribution of the paramagnetic probes previously incorporated in the outer leaflet of the plasma membrane was quantified as follows ( Bassé et al , 1992 ; Morrot et al , 1989 ). Spin‐labelled analogues (1–2% of endogenous PLs) were added to the platelet suspension at time zero from a concentrated solution in buffer B. Spin‐labelled PLs were incorporated into the outer plasma membrane leaflet in less than 1 min ( Seigneuret & Devaux, 1984).…”

Section: Methodsmentioning

confidence: 99%

Impaired redistribution of aminophospholipids with distinctive cell shape change during Ca²⁺‐induced activation of platelets from a patient with Scott syndrome

et al. 1998

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Summary.We have investigated phospholipid redistribution, membrane vesicle shedding, shape change, and granule release following A23187 activation of platelets from a patient with Scott syndrome, characterized by impaired transmembrane migration of phosphatidylserine (PS) accompanied by haemorrhagic complications, and two of her children. Electron spin resonance spectroscopy measurement of phospholipids redistribution showed that the internalization of PS was unaffected by the disorder but, after activation, PS exposure was significantly reduced in platelets from the homozygous-type patient. Vesicle shedding was also reduced in these platelets. However, the slow redistribution of phosphatidylcholine was similar to that observed in normal platelets. When treated with calpeptin, platelets from the homozygous-type patient, unlike normal or heterozygous Scott syndrome platelets, showed a smoothly rounded shape without filopods after activation. Following A23187 activation of normal platelets, filopod formation was consecutive to the re-exposition of aminophospholipids on the outer leaflet of the plasma membrane, and the existence of a floppase (outward aminoPLs translocase) has been suggested. In homozygous Scott syndrome platelets the deficiency in PS re-exposition, the absence of filopod formation, and low vesicle shedding are correlated with each other, and argue in favour of a disruption of the proposed floppase activity.

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“…Alternatively, prior egress of PS to the outer leaflet might create a mass imbalance that itself drives plasma membrane evagination and vesiculation (3,6,89,95). In this context, it has been observed that PS migration to the cell surface can precede membrane vesiculation, and can occur without microparticle formation (6,24,89,96). For instance, whereas calpain inhibitors have been observed to inhibit plasma membrane vesiculation, surface exposure of PS was not affected under these conditions (26,97).…”

Section: Relationship Of Ps Egress To Shedding Of Plasma Membrane Vesmentioning

confidence: 99%

Unraveling the Mysteries of Phospholipid Scrambling

Sims

Wiedmer²

2001

Thromb Haemost

167

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SummaryPlasma membrane phospholipid asymmetry is maintained by an aminophospholipid translocase that transports phosphatidylserine (PS) and phosphatidylethanolamine (PE) from outer to inner membrane leaflet. Cell activation or injury leads to redistribution of all major lipid classes within the plasma membrane, resulting in surface exposure of PS and PE. Cell surface-exposed PS can serve as receptor sites for coagulation enzyme complexes, and contributes to cell clearance by the reticuloendothelial system. The mechanism(s) by which this PL ”scrambling” occurs is poorly understood. A protein called phospholipid scramblase (PLSCR1) has been cloned that exhibits Ca2+-activated PL scrambling activity in vitro. PLSCR1 belongs to a new family of proteins with no apparent homology to other known proteins. PLSCR1 is palmitoylated and contains a potential protein kinase C phosphorylation site. It further contains multiple PxxP and PPxY motifs, representing potential binding motifs for SH3 and WW domains implicated in mediating protein-protein interactions. Although at least two proteins have been shown to associate with PLSCR1, the functional significance of such interaction remains to be elucidated. Evidence that PLSCR1 may serve functions other than its proposed activity as PL scramblase is also presented.

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Phospholipid transverse mobility modifications in plasma membranes of activated platelets: An ESR study

Cited by 10 publications

References 18 publications

Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Impaired redistribution of aminophospholipids with distinctive cell shape change during Ca²⁺‐induced activation of platelets from a patient with Scott syndrome

Unraveling the Mysteries of Phospholipid Scrambling

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Phospholipid transverse mobility modifications in plasma membranes of activated platelets: An ESR study

Cited by 10 publications

References 18 publications

Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Transverse Redistribution of Phospholipids during Human Platelet Activation: Evidence for a Vectorial Outflux Specific to Aminophospholipids

Impaired redistribution of aminophospholipids with distinctive cell shape change during Ca2+‐induced activation of platelets from a patient with Scott syndrome

Unraveling the Mysteries of Phospholipid Scrambling

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Impaired redistribution of aminophospholipids with distinctive cell shape change during Ca²⁺‐induced activation of platelets from a patient with Scott syndrome